Treatment of demyelinating disorders

ABSTRACT

This invention discloses methods and compositions for the treatment of demyelinating disorders. Specifically, the invention relates to the use guanabenz or guanabenz derivative for treating demyelinating disorders.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a national phase application under 35 U.S.C. § 371of International Application No. PCT/US2014/020896, filed Mar. 5, 2014,which claims the benefit of U.S. Provisional Application No. 61/772,875,filed on Mar. 5, 2013. The entire contents of each of theabove-referenced disclosures are specifically incorporated herein byreference without disclaimer.

STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

This invention was made with government support under NS034939, awardedby the National Institutes of Health. The government has certain rightsin the invention.

BACKGROUND OF THE INVENTION

Demyelinating disorders affect more than 2.5 million people worldwide. Ademyelinating disorder is a condition in which the myelin sheath of aneuronal cell is damaged. Myelin is an electrically insulating materialthat forms layers, or myelin sheaths, around neuronal axons. Myelinserves many functions and is essential for nervous system function. Oneof the central functions of myelin is to increase the speed ofelectrical impulses along an axon, another function is to preventelectrical current from dissipating from the axon. Myelin is produced bytwo cell types: oligodendrocytes and Schwann cells. Oligodendrocytes areglial cells that supply myelin to neurons of the central nervous system,Schwann cells provide myelin to peripheral neurons. Demyelinationresults in impaired transmission of neuronal impulses, resulting indiverse symptoms that possibly include motor impairments, sensoryimpairments, cognitive dysfunction, emotional disturbances, and impairedcoordination.

There are many demyelinating disorders, the most common of which ismultiple sclerosis. Multiple sclerosis (MS) is an autoimmune diseaseaffecting the brain and spinal cord, in which the body's own immunesystem attacks myelin or myelin producing cells. Although MS symptomsvary widely, one of the pathological hallmarks of MS is the presence ofmultiple demyelinated plaques in the CNS. Demyelinated plaques areassociated with blood-brain-barrier (BBB) breakdown, inflammation,demyelination, and axonal degeneration. BBB breakdown allows for theinfiltration of autoreactive cytokines that attack myelin and myelinassociated proteins such as myelin oligodendrocyte glycoprotein (MOG),proteolipid protein (PLP), or myelin basic protein (MBP), among others.In MS, early neurologic dysfunction is related to demyelination thatresults in slowed or blocked conductance of electrical signals. Mostsubjects suffering from MS exhibit a relapsing/remitting form of thedisease, which is characterized by discontinuous bouts of clinicalsymptoms (relapse) punctuated be periods of symptomatic relief(remission). Other subjects experience chronic, progressive MS, whereinsymptoms worsen over time without periods of symptomatic relief Somesubjects experience relapsing/remitting MS early in the course of thedisease but then eventually transition to a chronic, progressive form ofthe disease. Besides MS, other common demyelinating disorders includebut are not limited to acute disseminated encephalomyelitis,periventricular leukomalacia, periventricular white matter injury, TabesDorsalis, Devic's disease, optic neuritis, progressive multifocalleukoencephalopathy, transverse myelitis, chronic inflammatorydemyelinating polyneuropathy, anti-MAG peripheral neuropathy,adrenoleukodystrophy, adrenomyeloneuropathy, diffuse white matterinjury, Guillain-Barré Syndrome, central pontine myelinolysis, inheriteddemyelinating diseases such as leukodystrophy, and Charcot Marie ToothDisease. Sufferers of pernicious anemia or Vitamin B12 deficiency canalso suffer nerve damage if the condition is not diagnosed quickly.

The precise etiologies of many CNS disorders, including demyelinatingdisorders, are not fully characterized. Recent studies have implicatedendoplasmic reticulum (ER) stress response pathways in several CNSdisorders (J Cell Mol. Med. 2011 October; 15(10):2025-39). ER stress isa highly evolutionarily conserved pathway that serves to reduce theburden of misfolded or unfolded proteins in the ER. Even a modestaccumulation of unfolded/misfolded proteins in the ER can induce ERstress, compromising the ability of the ER to properly process secretoryand/or membrane proteins. The ER stress response pathway involves threemain signaling pathways, the IRE1 pathway, the ATF6 pathway, and thePERK pathway. Each of these pathways function to reduce ER burden andpromote cell survival, but may also ultimately result in cell death incases of persistent ER stress (Physiology 22:193-201, 2007). Inparticular, the PERK pathway has been implicated in demyelinatingdisorders such as MS. The PERK pathway leads to phosphorylation ofeukaryotic initiation factor 2-alpha (eIF2α), which has the main effectsof globally suppressing protein translation and selectively promotingtranslation of the transcription factor ATF4, which in turn activates anumber of target genes, including amino acid transporters and cellularredox genes (Physiology 22:193-201, 2007). The global suppression ofprotein translation serves to reduce the burden of misfolded/unfoldedproteins in the ER (Physiology 22:193-201, 2007).

Guanabenz is a small molecule guanidine derivative. It is known to be anα₂-adrenergic agonist, although it may also exert biological effectsthrough other mechanisms. Guanabenz is administered to treathypertension. The antihypertensive effects are thought to be mediated byits α₂-adrenergic agonist activity, resulting in reduced sympatheticstimulation of the heart, kidneys, and peripheral vasculature, andreduced blood pressure and pulse rate. Guanabenz is typicallyadministered twice a day as an oral tablet formulation. It has beenreported that guanabenz can impair memory function (Behav. Brain Res.2004 Aug. 31; 153(2):409-17) and reduce hippocampal neurogenesis (JNeurosci. 2010 Jan. 20; 30(3):1096-109). However, the use of guanabenzor its derivatives for the treatment of demyelinating disorders has notbeen reported. A study by Tsaytler et al (Science 2011 Apr. 1;332(6025):91-4) implies that guanabenz reduces ER stress burden, byreducing stress-induced dephosphorylation of eIF2α. However, this studyis limited to in vitro cell culture of epithelial cells. No in vivo datais provided, nor is there any hint for the in vivo use of guanabenz intreating any demyelinating disorders.

Presently, there are no known cures for demyelinating disorders,including multiple sclerosis. There are several drugs approved by theFDA for multiple sclerosis, however, these only provide some limitedsymptomatic relief. In fact, several of the currently approvedtreatments for demyelinating disorders are associated with significantadverse effects, such as flu-like symptoms, fever, liver damage, and insome cases, significantly increased risk for progressive multifocalleukoencephalopathy. Therefore, there is a pressing need for improvedtherapies for demyelinating disorders.

SUMMARY OF THE INVENTION

Accordingly, the present invention provides methods and compositionsthat address the limitations of current therapeutics for a variety ofdemyelinating disorders.

In one aspect, the present invention provides a method of increasingviability of a neuronal cell in a subject suffering from a demyelinatingdisorder, comprising administering to the subject suffering from ademyelinating disorder an effective amount of a compound of Formula I(below):

or derivative or pharmaceutically acceptable salt thereof, wherein R₁,R₂, R₃, R₄, and R₅ are independently hydrogen, deuterium, halogen,haloalkyl, alkyl, alkoxy, hydroxyl, aryl, or aryloxy, thereby increasingviability of a neuronal cell in said subject.

In another aspect, the present invention provides a method of treating asymptom of a demyelinating disorder, comprising the step ofadministering to a subject in need thereof an effective amount of acompound of Formula I, thereby treating a symptom of a demyelinatingdisorder in said subject.

In another aspect, the present invention provides a method of delayingonset of a symptom of a demyelinating disorder in a subject at risk ofthe demyelinating disorder, comprising administering to the subject atrisk of a demyelinating disorder an effective amount of a compound ofFormula I, thereby delaying onset of a symptom of a demyelinatingdisorder in the subject.

In another aspect, the present invention provides a method of reducingrelapse and/or severity of a symptom of a demyelinating disorder in asubject suffering from the demyelinating disorder, comprisingadministering to the subject at suffering from the demyelinatingdisorder an effective amount of a compound of Formula I, therebyreducing relapse and/or severity of a symptom of a demyelinatingdisorder in the subject.

In another aspect, the present invention provides a method of increasingviability of cells of the oligodendrocyte lineage when exposed to aninflammatory agent, comprising administering to the cells of theoligodendrocyte lineage with an amount of a compound of Formula I,effective in reducing apoptosis of said cells of the oligodendrocytelineage, wherein said apoptosis is induced by said inflammatory agent,thereby increasing viability of the cells of the oligodendrocytelineage.

In another aspect, the present invention provides a method of protectinga brain cell from inflammation in a subject in need thereof, the methodcomprising administering to the subject an effective amount of acompound of Formula I, thereby protecting a brain cell from inflammationin the subject. In some embodiments, the subject has or is suspected ofhaving amyotrophic lateral sclerosis (ALS).

In some aspects embodiments of the compound of Formula I describedherein, R₁ and R₅ are not hydrogen.

In some aspects embodiments of the compound of Formula I describedherein, R₁ and R₅ are halogen.

In some aspects embodiments of the compound of Formula I, the compoundis of Formula II (below) or derivative or pharmaceutically acceptablesalt thereof:

In some embodiments of the present invention, the demyelinating disorderis multiple sclerosis.

In other embodiments of the present invention, the demyelinatingdisorder is selected from the group consisting of: acute disseminatedencephalomyelitis, periventricular leukomalacia, periventricular whitematter injury, Tabes Dorsalis, Devic's disease, optic neuritis,progressive multifocal leukoencephalopathy, transverse myelitis, chronicinflammatory demyelinating polyneuropathy, anti-MAG peripheralneuropathy, adrenoleukodystrophy, adrenomyeloneuropathy, Guillain-BarréSyndrome, central pontine myelinolysis, diffuse white matter injury,inherited demyelinating diseases such as leukodystrophy, and CharcotMarie Tooth Disease.

In some embodiments, the symptom of a demyelinating disorder is selectedfrom the group consisting of: fatigue, somatosensory dysfunction,tingling, pain, numbness, balance problems, problems with walking,changes in vision, depression, emotional changes, mood swings, impairedcognition, muscle dysfunction, impaired muscle coordination, sexualimpairment, speech impairment, swallowing impairment, bladderdysfunction, bowel dysfunction.

In some embodiments of the invention, about 1 mg to about 64 mg of thecompound is administered to the subject.

In other embodiments of the invention, about 4 mg to about 20 mg of thecompound is administered to the subject.

In still yet others embodiment of the present invention, the compound isadministered orally.

In some embodiments, the brain cell is a neuronal cell. In otherembodiments, the brain cell is a glial cell. In still yet otherembodiments, the brain cell is a cell of the oligodendrocyte lineage.

In some embodiments, the neuronal cell is a neuron.

In some aspects of the invention, administration of said compoundsubstantially obviate relapse of a symptom of a demyelinating disorderin a subject over a course of at least 1 month.

In some embodiments of the invention, the compound of Formula I is acompound of Formula II, wherein upon administering an effective amountyields a reduction in apoptosis by at least 20%.

In other aspects of the invention, administration of said compoundreduces severity of a symptom of a demyelinating disorder in a subjectby at least about 50%.

In some aspects, the present invention provides an oral dosage formcomprising a unit dosage of guanabenz present in an amount effective inameliorating a symptom of a demyelinating disorder, wherein the oraldosage form comprises instructions for use of said dosage form for asubject suffering from said demyelinating disorder. In some embodimentsof the oral dosage form, the instructions specify administering the unitdosage according to a regime having at least one dose per day.

In some cases, this disclosure provides an oral dosage form comprisingabout 64 mg of a compound provided herein, such as guanabenz.

In some cases, administration of a compound provided in this disclosureis combined with at least one another agent. In some embodiments, the atleast one other agent is useful for the treatment of a demyelinatingdisorder. In some embodiments, the at least one other agent isglatiramer acetate. In some cases, guanabenz and glatiramer acetate areboth administered to a subject. In some cases, about 64 mg of guanabenzis administered to a subject receiving glatiramer acetate. In someembodiments, the at least one other agent is interferon-beta. In someembodiments, the at least one other agent is interferon beta-1a. In somecases, guanabenz and interferon beta-1a are both administered to asubject. In some cases, about 64 mg of guanabenz is administered to asubject receiving interferon beta-1a. In some embodiments, the at leastone other agent is interferon beta-1b. In some cases, guanabenz andinterferon beta-1b are both administered to a subject. In some cases,about 64 mg of guanabenz is administered to a subject receivinginterferon beta-1b. In some embodiments, the at least one other agent isdimethyl fumerate (BG-12). In some cases, guanabenz and dimethylfumerate are both administered to a subject. In some cases, about 64 mgof guanabenz is administered to a subject receiving dimethyl fumerate.In some embodiments, the at least one other agent is fingolimod(FTY720). In some cases, guanabenz and fingolimod are both administeredto a subject. In some cases, about 64 mg of guanabenz is administered toa subject receiving fingolimod. In some embodiments, the at least oneother agent is mitoxantrone. In some cases, guanabenz and mitoxantroneare both administered to a subject. In some cases, about 64 mg ofguanabenz is administered to a subject receiving mitoxantrone. In someembodiments, the at least one other agent is natalizumab. In some cases,guanabenz and natalizumab are both administered to a subject. In somecases, about 64 mg of guanabenz is administered to a subject receivingnatalizumab. In some embodiments, the at least one other agent isdalfampridine. In some cases, guanabenz and dalfampridine are bothadministered to a subject. In some cases, about 64 mg of guanabenz isadministered to a subject receiving dalfampridine. In some embodiments,the at least one other agent is teriflunomide. In some cases, guanabenzand teriflunomide are both administered to a subject. In some cases,about 64 mg of guanabenz is administered to a subject receivingteriflunomide. In some embodiments, the at least one other agent isdaclizumab. In some cases, guanabenz and daclizumab are bothadministered to a subject. In some cases, about 64 mg of guanabenz isadministered to a subject receiving daclizumab.

In some cases, the 64 mg of guanabenz is administered in a single dose.In some cases, the single dose is administered to the subject in theevening.

In some cases, this disclosure provides a composition comprising acompound provided herein and an additional therapeutic agent. In someembodiments, the additional therapeutic agent is an agent useful for thetreatment of a demyelinating disorder. In some embodiments, theadditional therapeutic agent is glatiramer acetate. In some embodiments,the additional therapeutic agent is dimethyl fumerate (BG-12). In someembodiments, the additional therapeutic agent is dimethyl fumerate(BG-12). In some embodiments, the additional therapeutic agent isfingolimod (FTY720). In some embodiments, the additional therapeuticagent is interferon beta-1a. In some embodiments, the additionaltherapeutic agent is interferon beta-1b. In some embodiments, theadditional therapeutic agent is mitoxantrone. In some embodiments, theadditional therapeutic agent is natalizumab. In some embodiments, theadditional therapeutic agent is dalfampridine. In some embodiments, theadditional therapeutic agent is teriflunomide. In some embodiments, theadditional therapeutic agent is daclizumab. In some cases, the compoundis guanabenz. In some cases, the composition comprises apharmaceutically acceptable carrier. In some cases, the pharmaceuticallyacceptable carrier is suitable for injection. In some cases, theinjection is a subcutaneous injection. In some cases, the injection isintravenous administration.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 illustrates the canonical ER stress response pathway. The red boxindicates the pathway modulated by Guanabenz, resulting in decreasedGADD34-PP1 activity and reduced eIF2α dephosphorylation.

FIG. 2A depicts photomicrographs of a fluorescein-diacetate/propidiumiodide assay for cell viability in differentiating rat oligodendrocyteprecursor cells (drOPCs) treated with interferon-gamma (IFN-γ) alone orin combination with guanabenz, where green cells are viable cells andred cells are non-viable cells.

FIG. 2B illustrates the quantitation of the results shown in FIG. 2A.

FIG. 3A depicts photomicrographs of a TUNEL assay for apoptosis indrOPCs treated with interferon-gamma (IFN-γ), alone or in combinationwith guanabenz.

FIG. 3B illustrates the quantitation of the results shown in FIG. 3A.

FIG. 4 depicts cultured rat cerebellar slices stained for myelin basicprotein and toluidine blue. The depicted rat cerebellar samples weretreated with IFN-γ alone or in combination with 2.5-10 μM guanabenz.

FIG. 5A depicts clinical scores over the course of experimentalautoimmune encephalitis (EAE), a mouse model of chronic multiplesclerosis. The mice were treated with 4-16 mg/kg/day of guanabenz orwith control vehicle.

FIG. 5B depicts average age of symptom onset over the course ofexperimental autoimmune encephalitis (EAE), a mouse model of chronicmultiple sclerosis. The mice were treated with 4-16 mg/kg/day ofguanabenz or with control vehicle.

FIG. 6 depicts a plot of clinical scores over days post symptom onset.The mice were treated with 4-16 mg/kg/day of guanabenz or with controlvehicle.

FIG. 7A depicts clinical scores over the course of remitting/relapsingEAE, a mouse model of remitting/relapsing multiple sclerosis. The micewere treated with 8 mg/kg/day guanabenz or control vehicle.

FIG. 7B depicts clinical scores over the course of remitting/relapsingEAE, a mouse model of remitting/relapsing multiple sclerosis. The micewere treated with 16 mg/kg/day guanabenz or control vehicle.

FIG. 8 depicts the time course of serum and brain concentrations ofguanabenz, expressed as ng/ml or ng/g, in mice administered 4-16 mg/kgguanabenz daily after EAE induction.

FIG. 9 depicts the time course of serum and brain concentrations ofguanabenz, expressed in μM concentrations, in mice administered 4-16mg/kg guanabenz daily after EAE induction.

FIG. 10 depicts inflammatory ear-swelling response to a local injectionof MOG₃₅₋₅₅, the antigen used to induce EAE. The mice were treated with8 mg/kg/day guanabenz or control vehicle.

FIG. 11 depicts effects of guanabenz/interferon co-administration onclinical symptoms in chronic EAE mice. The mice were treated withcontrol vehicle, 8 mg/kg/day guanabenz, 5000 U/day interferon, or acombination of guanabenz and interferon.

DETAILED DESCRIPTION OF THE INVENTION

Overview

In general, described below are methods and compositions for the use ofguanabenz or guanabenz derivatives for the treatment of demyelinatingdisorders, including but not limited to multiple sclerosis. Disclosedherein are a number of methods for the use of guanabenz or guanabenzderivatives for the treatment of, for ameliorating the symptoms of, fordelaying onset of symptoms of, or for reducing relapse severity ofdemyelinating disorders. Also disclosed are methods for the use ofguanabenz or guanabenz derivatives to provide neuroprotection to asubject in need thereof. In some embodiments the subject is sufferingfrom a demyelinating disorder. In some embodiments the subject issuffering from or is suspected of having Amyotrophic lateral sclerosis(ALS). Furthermore, disclosed herein are methods for the use ofguanabenz or guanabenz derivatives to provide protection of cells of theoligodendrocyte lineage, the cells providing the myelin sheath in thecentral nervous system, wherein the cells of the oligodendrocyte lineagemay suffer damage, stress, or death due to inflammatory insult. Alsodisclosed is a composition for an oral dosing package, comprisinginstructions for use of said dosage form for a subject suffering from ademyelinating disorder.

DEFINITIONS

As used in the specification and claims, the singular form “a”, “an” and“the” include plural references unless the context clearly dictatesotherwise. For example, the term “a symptom” may include a plurality ofsymptoms.

The term “about” means within an acceptable error range for theparticular value as determined by one of ordinary skill in the art. Insome cases, the acceptable error range may depend in part on thelimitations of the measurement system. For example, “about” can meanwithin 1 or more than 1 standard deviation, per the practice in the art.Alternatively, “about” can mean a range of up to 25%, preferably up to10%, more preferably up to 5%, and more preferably still up to 1% of agiven value. Alternatively, the term can mean within an order ofmagnitude, preferably within 10-fold, preferably within 5-fold, and morepreferably within 2-fold, of a value. Where particular values aredescribed in the application and claims, unless otherwise stated theterm “about” meaning within an acceptable error range for the particularvalue should be assumed.

For the purposes of this invention, a “demyelinating disorder” is anycondition involving dysfunction of the myelin sheath encasing neuronalaxons. Examples of demyelinating disorders include but are not limitedto multiple sclerosis, acute disseminated encephalomyelitis,periventricular leukomalacia, periventricular white matter injury, TabesDorsalis, Devic's disease, Optic neuritis, progressive multifocalleukoencephalopathy, transverse myelitis, chronic inflammatorydemyelinating polyneuropathy, anti-MAG peripheral neuropathy,Adrenoleukodystrophy, adrenomyeloneuropathy, Guillain-Barré Syndrome,central pontine myelinolysis, diffuse white matter injury, inheriteddemyelinating diseases such as demyelinating leukodystrophies, andCharcot Marie Tooth Disease. In some embodiments, the demyelinatingdisorder can be the result of pernicious anemia or vitamin B12deficiency.

The terms “treating” or “treating a symptom of” a demyelinating disorderare used interchangeably. These terms refer to, but are not limited to,utilizing a method or methods to achieve a therapeutic benefit and/or aprophylactic benefit. In some cases, a therapeutic benefit may refer toreducing the severity of a symptom or symptoms of a demyelinatingdisorder. In some cases, a therapeutic benefit may refer to addressingor correcting the biological mechanisms underlying the demyelinatingdisorder. In other cases, a therapeutic benefit may refer to halting orslowing the progression of a demyelinating disorder. In other cases, atherapeutic benefit may refer to reducing demyelination or to promotingremyelination. In yet other cases, a therapeutic benefit may refer toreducing inflammation associated with a number of demyelinatingdisorders. In yet other cases, a therapeutic benefit may refer toprotecting brain cells from inflammation associated with a number ofdemyelinating disorders. In yet other cases, a therapeutic benefit mayrefer to reducing the size and/or number of white matter plaques and/orlesions. In still yet other cases, a therapeutic benefit may refer toincreasing the viability and/or health of cells of the oligodendrocytelineage. In still yet other cases, therapeutic benefit may refer toproviding neuroprotection of axons. In addition, a prophylactic benefitmay refer to reducing the risk of developing a demyelinating disorder.In some cases, a prophylactic benefit may refer to delaying the onset ofsymptom(s) of a demyelinating disorder. In some cases, a prophylacticbenefit may refer to reducing or delaying relapse of a demyelinatingdisorder.

In the present invention, “the treatment” can involve administering to asubject a therapeutically effective amount of guanabenz or a guanabenzderivative. In some embodiments, the composition comprises apharmaceutically acceptable carrier. In some embodiments, theadministration of guanabenz or guanabenz derivative may involveco-administration with another therapeutic agent for the treatment of ademyelinating disorder, including, but not limited to, glatirameracetate, dimethyl fumerate (BG-12), fingolimod (FTY720), interferonbeta-1a, interferon beta-1b, mitoxantrone, natalizumab, dalfampridine,teriflunomide, or daclizumab.

The terms “co-administration,” “administered in combination with”,“combination therapy” and their grammatical equivalents, encompassadministration of two or more agents, one of which comprises thecompound(s) disclosed in the present invention, to a subject so that theagents and/or their metabolites are present in the animal at the sametime. Co-administration may involve simultaneous administration inseparate compositions, or may involve administration at different timesin separate compositions, or may involve administration in a singlecomposition in which the agents are present. Co-administered agents maybe in the same formulation. Co-administered agents may also be indifferent formulations.

“An effective amount” and “a therapeutically effective amount” are usedinterchangeably. “An effective amount” is an amount of the compositionthat achieves a therapeutic benefit and/or prophylactic benefit. Thetherapeutically effective amount can be determined by a physician or oneof ordinary skill in the art. One of ordinary skill in the art willunderstand that an effective amount may vary from subject to subject,depending on the subject's size, age, general health, route ofadministration, the subject's level of risk for developing ademyelinating disorder, the subject's severity of symptom(s), thesubject's progression of the disorder, or other factors. One of ordinaryskill in the art will understand that an effective amount for aparticular subject may change over time, depending on factors describedabove.

A “sub-therapeutic amount” of an agent is an amount less than theeffective amount for that agent. When combined with an effective orsub-therapeutic amount of one or more additional agents, thesub-therapeutic amount can produce a result desired by the physician,due to, for example, synergy in the resulting efficacious effects, orreduced adverse effects.

A “synergistically effective” therapeutic amount or “synergisticallyeffective” amount of an agent or therapy is an amount which, whencombined with an effective or sub-therapeutic amount of one or moreadditional agents, produces a greater effect than when either of theagents are used alone. In some embodiments, a synergistically effectivetherapeutic amount of an agent or therapy produces a greater effect whenused in combination than the additive effects of any of the individualagents when used alone. The term “greater effect” encompasses not only areduction in symptoms of the disorder to be treated, but also animproved side effect profile, improved tolerability, improved patientcompliance, improved efficacy, or any other improved clinical outcome.

As used herein, “agent” or “biologically active agent” refers to abiological, pharmaceutical, or chemical compound or other moiety.Non-limiting examples include a simple or complex organic or inorganicmolecule, a peptide, a protein, an oligonucleotide, an antibody, anantibody derivative, an antibody fragment, a vitamin or vitaminderivative, a carbohydrate, a toxin, a chemotherapeutic compound, or avaccine. Various compounds can be synthesized, for example, smallmolecules and oligomers (e.g., oligopeptides and oligonucleotides), andsynthetic organic compounds based on various core structures. Inaddition, various natural sources can provide compounds for screening,such as plant or animal extracts, and the like. One of skill in the artcan readily recognize that there is no limit as to the structural natureof the agents of the present invention.

In some embodiments, “the subject” is an animal. In some embodiments,the subject is a human being.

The term “pharmaceutically acceptable salt” refers to salts derived froma variety of organic and inorganic counter ions well known in the art.Pharmaceutically acceptable salts that will be suitable for guanabenzand guanabenz derivatives include, but are not limited to, hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andacetic acid.

The term “pharmaceutically acceptable carrier” refers to a solid orliquid filler, diluents, or encapsulating substance, and may include anyand all solvents, media, coatings and the like. The use of such mediaand agents for pharmaceutically active substances is well known in theart. Other active ingredients may also be incorporated into thecompositions.

The term “brain cell” may refer to any cell type found in the brain.Examples of brain cells include but are not necessarily limited toneuronal cells, neurons, glial cells, cells of the oligodendrocytelineage, astrocytes, microglia, endothelial cells. Characteristics ofthese cell types are well known to those skilled in the art.

The term “a neuronal cell” refers to a cell of neuronal lineage.Exemplary cells include but are not limited to neurons, neuralprecursors, glial cells, cells of the oligodendrocyte lineage,astrocytes.

An “alkyl” group refers to a saturated aliphatic hydrocarbon group. Thealkyl moiety, whether saturated or unsaturated, may be branched,straight chain, or cyclic. An “alkyl” moiety may, for example, have 1 to10 carbon atoms (whenever it appears herein, a numerical range such as“1 to 10” refers to each integer in the given range; e.g., “1 to 10carbon atoms” means that the alkyl group may consist of 1 carbon atom, 2carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms,although the present definition also covers the occurrence of the term“alkyl” where no numerical range is designated). The alkyl group couldalso be a “lower alkyl” having 1 to 5 carbon atoms. The alkyl group ofthe compounds described herein may be designated as “C1-C4 alkyl”,“C5-C8 alkyl” or similar designations. By way of example only, “C1-C4alkyl” indicates that there are one to four carbon atoms in the alkylchain, i.e., the alkyl chain is selected from the group consisting ofmethyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, andt-butyl. Common alkyl groups include, but are in no way limited to,methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl,pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and thelike.

The term “alkoxy” as used herein means an —O-alkyl group, wherein alkylis as defined above. Alkoxy groups include moieties such as, but notlimited to, methoxy, ethoxy, n-propyloxy, i-propyloxy, n-butyloxy,i-butyloxy, t-butyloxy, pentyloxy, hexyloxy, including their isomers.

The term “aryl” as used herein refers to an aromatic carbocyclic systemof about 6 to 14 carbon atoms, which can include a single ring ormultiple aromatic rings fused or linked together where at least one partof the fused or linked rings forms the conjugated aromatic system. Thearyl groups include, but are not limited to, phenyl, naphthyl, biphenyl,anthryl, tetrahydronaphthyl, phenanthryl, indene, benzonaphthol andfluorenyl.

The term “aryloxy” as used herein denotes a O-aryl group, wherein arylis as defined above. An aryloxy group can be unsubstituted orsubstituted with one or two suitable substituents.

The term “halogen” or “halo” as used herein denotes fluorine, chlorine,bromine, or iodine.

The term “haloalkyl” as used herein denotes a unbranched or branchedchain alkyl group as defined above wherein 1, 2, 3 or more hydrogenatoms are substituted by a halogen. Examples include, but are notlimited to, 1-fluoromethyl, 1-chloromethyl, 1-bromomethyl, 1-iodomethyl,trifluoromethyl, trichloromethyl, tribromomethyl, triiodomethyl,1-fluoroethyl, 1-chloroethyl, 1-bromoethyl, 1-iodoethyl, 2-fluoroethyl,2-chloroethyl, 2-bromoethyl, 2-iodoethyl, 2,2-dichloroethyl,3-bromopropyl or 2,2,2-trifluoroethyl.

Subjects in Need of Treatment

In one aspect of the present invention, the treatment is administered toa subject suffering from a demyelinating disorder. In some cases, asubject suffering from a demyelinating disorder can be a subjectdiagnosed, suspected of having, experiencing or having experienced oneor more symptom(s) of a demyelinating disorder. The symptom(s) of ademyelinating disorder include but are not limited to fatigue,somatosensory dysfunction, tingling, pain, numbness, balance problems,problems with walking, changes in vision, depression, emotional changes,mood swings, impaired cognition, muscle dysfunction, impaired musclecoordination, sexual impairment, speech impairment, swallowingimpairment, bladder dysfunction, bowel dysfunction. One of ordinaryskill in the art will understand that the symptom(s) of demyelinatingdisorders may vary widely from subject to subject, in terms of symptommanifestation, onset, and severity. In some cases, the symptom(s) maylast for 24 hours and then subside. In some cases after a subject'ssymptom(s) have subsided, the subject may experience any of thesymptom(s) at a later time (relapsing/remitting disorder). In some casesthe symptom(s) may progressively worsen over time (progressivedisorder).

In some cases, a subject may be diagnosed with or suspected of having ademyelinating disorder based on any combination of a number of tests andassays described below. One of ordinary skill in the art will understandthat the tests and assays described below may also be used to monitorthe progression of the disorder or to monitor symptom severity orsymptom relapse. The term “diagnosis” as used herein encompasses themonitoring of the disorder and/or its symptom severity or relapse.

In some cases, diagnosis of the demyelinating disorder can be based on aphysical exam, neurological exam, medical history, electrophysiologicaltest, lab assay, or imaging. In some cases, a physical exam or medicalhistory will include an assessment or history of symptoms. In somecases, the neurological exam will include an assessment of the patient'svision, reflexes, balance, coordination, and/or muscle strength.

In some cases, the electrophysiological test can involve examining theelectrical impulses traveling through the nerves to determine if theimpulses are moving normally or too slowly. In some cases, theelectrophysiological test can involve placing wires on the scalp to testthe subject's evoked potential response to certain types of stimulation.In some cases, the stimulation may be visual in nature. In other cases,the stimulation may be auditory in nature. In yet other cases, thestimulation may be somatosensory in nature. In still yet other cases,the stimulation may involve stimulation of muscle responses in theleg(s) or arm(s). One of ordinary skill in the art will understand thatthe measurement of normal speed evoked potentials in a subject does notnecessarily preclude a diagnosis of a demyelinating disorder.

In some cases, the lab assay may include a cerebrospinal fluid (CSF)assessment of biomarkers associated with a demyelinating disorder. Insome cases, CSF assessment may involve an assessment of immunoglobulinlevels in the cerebrospinal fluid. It is known in the art that anincrease in immunoglobulin concentrations in CSF is associated withdemyelinating disorders. In some embodiments, the assessment ofimmunoglobulin may involve taking an IgG index (a comparison between IgGlevels in the CSF and in blood serum). In some cases, an elevated IgGindex can be used to support diagnosis of a demyelinating disorder. Insome embodiments, assessment of CSF may involve separation of CSFproteins by electrophoresis and identification of oligoclonalimmunoglobulin bands by electrophoresis. In some cases, oligoclonalbands from CSF appear as strips on the electrophoresis gel, and may bevisualized by Coomassie Blue staining or other staining technique. Insome cases, the presence of oligoclonal immunoglobulin in CSF may beused to support diagnosis of a demyelinating disorder. In someembodiments, assessment of CSF may involve determination of overallprotein levels in the CSF sample. In some cases, elevated protein levelsin the CSF may be used to support diagnosis of a demyelinating disorder.In some cases, elevated CSF protein levels may be used to supportdiagnosis of relapse of a demyelinating disorder. In some embodiments,assessment of CSF may involve measuring a cell count of the CSF sample.In some embodiments, a higher cell count in the CSF sample can be usedto support diagnosis of a demyelinating disorder. In some cases, thecells found in CSF may be T-lymphocytes. In some cases, the presence ofT-lymphocytes in CSF may be used to support diagnosis of a demyelinatingdisorder.

In some embodiments, the lab assay may involve testing for the presenceof autoantibodies in a biological sample from the subject. In someembodiments, the sample is a blood serum sample. In some embodiments,the autoantibodies to be assayed are directed to myelin or amyelin-associated protein. In some embodiments, the autoantibody to beassayed is a myelin basic protein antibody. In some embodiments, theautoantibody to be assayed is a myelin oligodendrocyte glycoproteinantibody. In some embodiments, the assay to detect autoantibodies is awestern blot assay.

In some cases, imaging can involve magnetic resonance imaging (MRI) ofmyelin in the central nervous system of a subject, comprising thesubject's brain and spinal cord. In some cases, magnetic resonanceimaging can involve intravenous injection of gadolinium-based tracers tovisualize the white matter. In some cases, magnetic resonance imagingcan be used to identify white matter plaques and/or lesions associatedwith the disease. In some cases, white matter plaques and/or lesionsexhibit high intensity on T2-weighted and FLAIR images and low intensityon T1-weighted scans. One of ordinary skill in the art will realize thatintensities of white matter plaques and/or lesions will vary dependingon magnetic field strength, pulse sequence parameters, and partialvolume effects. In some cases, the white mater plaques and/or lesionsmay show partial or complete destruction of myelin (demyelination). Insome cases, white matter plaques and/or lesions may occur in the brain.In some cases, white matter plaques and/or lesions may occur in thespinal cord. In some cases, the diagnosis is based on the presence oftwo or more white matter plaques and/or lesions. In some cases, thedemyelination may occur in a perivenular distribution. In some cases,the demyelination may be associated with gliosis. In some cases, thedemyelination may be associated with hallmarks of inflammation. In somecases, the hallmarks of inflammation may include infiltration ofmononuclear cells and/or lymphocytes. One of ordinary skill in the artwill understand that the lack of white matter plaques and/or lesionsvisualized by imaging does not necessarily preclude a diagnosis of ademyelinating disorder, as some lesions may be too small to bevisualized by imaging.

In some cases, the symptom(s) of a demyelinating disorder may overlapwith a number of other diseases or syndromes. In some cases, diagnosisof a demyelinating disorder may involve ruling out other possibleconditions that may cause similar symptom(s). In some cases, conditionsthat may be ruled out to support a diagnosis of a demyelinating disorderinclude, but are not limited to, stroke, alcoholism, emotionaldisorders, Lyme disease, chronic fatigue syndrome, fibromyalgia, AIDS,cervical spondylosis, hyperthyroidism, scleroderma, Sjogren's syndrome,systematic lupus erythematosus. Methods for ruling out the abovedisorders are known to those skilled in the art.

In another aspect of the present invention, the treatment isadministered to a subject at increased risk for developing ademyelinating disorder. A number of environmental and genetic factorshave been found to be associated with increased risk for demyelinatingdisorders. In some embodiments of the invention, a subject may beconsidered to have an increased risk for developing a demyelinatingdisorder based on any combination of the risk factors described herein.One of ordinary skill in the art will understand that a greater numberof risk factors that can be applied to a subject may correlate withgreater risk of the subject developing a demyelinating disease, however,a subject may be considered to be at increased risk for developing ademyelinating disorder based on even one of the risk factors describedherein.

In some embodiments, the subject may be considered to be at increasedrisk for developing a demyelinating disorder if the subject isexperiencing or has experienced a clinically isolated syndrome, whereinthe clinically isolated syndrome is a first or isolated neurologicepisode that lasts at least 24 hours, wherein the subject experiences atleast one or more symptom(s) described herein. The clinically isolatedsyndrome may be monofocal, wherein the subject experiences a singleneurologic symptom described herein. The clinically isolated syndromemay be multifocal, wherein the subject experiences more than one symptomdescribed herein. In some cases, the clinically isolated syndrome may beassociated with inflammation and/or demyelination in one or more sitesin the central nervous system.

In some embodiments, the subject may be considered to be at increasedrisk for developing a demyelinating disorder if the subject exhibits oneor more MRI white matter plaques and/or lesions without any discernableclinical symptoms. Methods for assessing white matter plaques and/orlesions are described herein.

In some embodiments, the subject may be considered to be at increasedrisk for developing a demyelinating disorder if the subject exhibits CSFbiomarkers associated with a demyelinating disorder. CSF biomarkersassociated with a demyelinating disorder have been described herein.

In some embodiments, a subject may be considered to be at increased riskfor developing a demyelinating disorder if the subject has a familyhistory of a demyelinating disorder. In some embodiments, a familyhistory of a demyelinating disorder may involve any relative of thesubject being diagnosed or suspected of having a demyelinating disorder.In some embodiments, a family history of a demyelinating disorder mayinclude an immediate family member having been diagnosed or suspected ofhaving a demyelinating disorder. In some cases, the family memberdiagnosed or suspected of having a demyelinating disorder may be aparent. In some cases, the family member may be a sibling. In somecases, the sibling may be an identical twin. In some cases, the familymember may be a son or daughter. In some cases, the relative of thesubject can be outside the immediate family, e.g., a first cousin, asecond cousin, a third cousin, an aunt, an uncle, a second or thirdaunt, a second or third uncle, a niece, a nephew, a second or thirdniece, a second or third nephew.

In some embodiments, a subject may be considered to be at increased riskfor developing a demyelinating disorder if the subject harbors aparticular allele, polymorphism, or haplotype associated with ademyelinating disorder. Methods for determining whether a subjectharbors an allele, polymorphism, or haplotype are known to those ofordinary skill in the art, and may include but are not limited togenotyping methods, exome or genome sequencing methods, or haplotypeanalysis. In some embodiments, the allele, polymorphism, or haplotypeinvolves the Major Histocompatibility Complex class 2 (MHC II),otherwise known as human leukocyte antigen (HLA). In some cases, theallele is the HLA-DR2 allele, which has previously been associated withmultiple sclerosis. In some cases, the allele is the DRB1*1501 allele.In some cases, the allele is the DQA1*0102 allele. In some cases, theallele is the DQB1*0602 allele. In some cases, the haplotype is the HLAclass II DR15 haplotype (DRB1*1501-DQA1*0102-DQB1*0602). In someembodiments, the polymorphism may be a single nucleotide polymorphism(SNP) in a gene encoding an interleukin receptor. In some embodiments,the interleukin receptor may be interleukin 2 receptor-alpha(IL2R-alpha). In some embodiments, the IL2R-alpha SNP may be rs12722489.In some embodiments, the IL2R-alpha SNP may be rs2104286. In someembodiments, the interleukin receptor may be interleukin 7receptor-alpha (IL7R-alpha). In some embodiments, the IL7R-alpha SNP maybe Rs6897932. In some embodiments, the IL7R-alpha SNP may cause areduction in the amount of IL7R-alpha protein present at the surface ofT-cells. One of ordinary skill in the art will understand that the abovelist may not be a complete list of genetic risk factors fordemyelinating disorders.

In some embodiments, the subject may be considered to be at increasedrisk for developing a demyelinating disorder if the subject or arelative of the subject suffers or has suffered from another autoimmunedisorder. In some cases, the subject or relative may suffer or havesuffered from irritable bowel syndrome. In some cases, the subject orrelative may suffer or have suffered from autoimmune thyroid disease. Insome cases, the subject or relative may suffer or have suffered fromrheumatoid arthritis. In some cases, the subject or relative may sufferor have suffered from Crohn's Disease. In some cases, the subject orrelative may suffer or have suffered from Type 1 Diabetes.

In some embodiments, the subject may be considered to be at increasedrisk for developing a demyelinating disorder if the subject hasexperienced a disruption of the blood-brain barrier. Possible causes ofblood-brain barrier disruption include but are not limited to, traumaticbrain injury, viral infection, HIV infection, inflammation of themeninges, stroke, among others. In some embodiments, blood brain barrierdisruption may be indicated by the detection of serum markers in CSF. Insome embodiments, the serum markers may include: serum S-100beta. Insome embodiments, blood-brain barrier disruption may be indicated by thepresence of one or more white matter plaques and/or lesions asvisualized by MRI. Methods for visualizing white matter plaques and/orlesions are described herein.

In some embodiments, a subject may be considered to be at increased riskfor developing a demyelinating disorder if the subject is female.

In some embodiments, a subject may be considered to be at increased riskfor developing a demyelinating disorder if the subject is between 15 and60 years of age, or between 20 and 50 years of age, or around the age of20-30.

In some embodiments, a subject may be considered to be at increased riskfor developing a demyelinating disorder if the subject is of Caucasiandescent. In some embodiments, a subject may be considered to be atincreased risk for developing a demyelinating disorder if the subject isof northern European descent. In some embodiments, a subject may beconsidered to be at increased risk for developing a demyelinatingdisorder if the subject is of Scottish descent.

In some embodiments, a subject may be considered to be at increased riskfor developing a demyelinating disorder if the subject has experienced aviral infection. In some embodiments, the viral infection may be from ahuman herpesvirus. In some embodiments, the human herpesvirus may beEpstein-Barr virus. In some embodiments, the human herpesvirus may behuman herpesvirus 6 (HHV-6).

In some embodiments, the subject may be considered to be at increasedrisk for a demyelinating disease if the subject has had insufficientexposure to sunlight.

In some embodiments, the subject may be considered to be at increasedrisk for a demyelinating disease if the subject has a Vitamin Ddeficiency. In some cases, the subject may be absorbing or ingestingless than 1000 U of Vitamin D per day.

In some embodiments, the subject may be considered to be at increasedrisk for a demyelinating disease if the subject has ever been exposed tocigarette smoke.

Uses of Guanabenz or Guanabenz Derivatives

The present invention provides the use of a compound of Formula I(below):

or derivative thereof, or a pharmaceutically acceptable salt thereof,wherein R₁, R₂, R₃, R₄, and R₅ are independently hydrogen, deuterium,halogen, haloalkyl, alkyl, alkoxy, hydroxyl, aryl, or aryloxy, for thetreatment of a variety of demyelinating disorders. In a preferredembodiment, the compound may be a compound of Formula II (below):

herein referred to as “guanabenz” or a pharmaceutically acceptable saltthereof.

In some aspects of the present invention, a compound of Formula I orFormula II may be used to increase viability of a neuronal cell or cellof the oligodendrocyte lineage in a subject suffering from ademyelinating disorder. One of skill in the art may use various assays,tests, diagnostic methods, or indicators to determine whetheradministration of a compound of Formula I or Formula II increases theviability of a neuronal cell. In some embodiments, one may determine ifa compound of Formula I or Formula II has improved viability of aneuronal cell based on a cell culture assay. Examples of useful cellculture assays include but are not limited to counting the number ofcells in a cell population that has been contacted with a compound ofFormula I or Formula II for comparison to the number of cells in a cellpopulation that has not been contacted with guanabenz, measuringviability by a LIVE/DEAD® assay (Invitrogen), measuring cell death orapoptosis by a TUNEL assay. In some embodiments, one may determine if acompound of Formula I or Formula II has improved viability of a neuronalcell based on an ex vivo assay. Examples of ex vivo assays include butare not limited to, staining ex vivo tissue samples with cresyl violetand counting the number of stained cells, staining ex vivo tissuesamples using a TUNEL assay, staining ex vivo tissue samples withmarkers of brain cells or neuronal cells and counting the number ofcells containing the marker. Examples of useful markers include but arenot limited to, NeuN (a neuron-specific marker), GFAP (anastrocyte-specific marker), oligodendrocyte lineage specific markerssuch as oligodendrocyte specific protein, oligodendrocyte marker O1,oligodendrocyte marker O4, etc. In some embodiments, one may determineif a compound of Formula I or Formula II has improved viability of aneuronal cell based on non invasive, in vivo assays. In someembodiments, in vivo imaging of neurodegeneration may be used toindicate viability, wherein reduction of neurodegeneration is indicativeof increased viability. In some embodiments, neurodegeneration in vivomay be indicated by reduction in white or gray matter volume asdetermined by MRI, reduction in the number or integrity of axons asvisualized by diffusion tensor imaging, or by the appearance of plaquesor lesions in the brain. In some embodiments, methods for thenon-invasive, in vivo assessment of apoptosis may be used to determineviability, wherein reduction of apoptosis is indicative of increasedviability. Methods for the non-invasive, in vivo assessment of apoptosisare in development, for example, a radiolabeled analog is currentlybeing evaluated as a PET probe in a phase II study (Adv. Drug Deliv Rev2005; 57: 1087-1108, herein incorporated by reference). In someembodiments, increased viability of neuronal cells or cells of theoligodendrocyte lineage may also result in symptomatic improvement,e.g., improvement or delayed onset of any of the symptom(s) listedherein.

In some embodiments, administration of a compound of Formula I orFormula II may result in an increase in viability of about 0.1%, 0.5%,1%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or morethan 100%. In some embodiments, a compound of Formula I or Formula IImay increase viability by a range of about 0.1%-10%, about 0.5%-20%,about 1%-30%, about 5%-40%, about 10%-60%, about 20%-80%, about 30%-90%,about 40% to more than 100%.

Protecting Brain Cells from Inflammation

In some aspects of the present invention, a compound of Formula I orFormula II may be used to protect brain cells from inflammation. One ofskill in the art may use various assays, tests, diagnostic methods, orindicators to determine whether a subject is in need of a compound ofFormula I or Formula II for the protection of brain cells frominflammation, or to determine if a cell of the oligodendrocyte lineageof a subject has likely been exposed to an inflammatory agent. In somecases, a subject may be determined to be in need of protection frominflammation or exposed to an inflammatory agent based on CSF assaysdescribed herein, or by PET imaging of [¹¹C](R)-PK11195, which binds toactivated microglia. In some cases, subjects that harbor one or morehaplotypes or genetic variants associated with demyelinating diseasesmay be considered to likely harbor neuroinflammation, and thus may beconsidered subjects in need of a compound of Formula I or Formula II forthe protection of brain cells against inflammation. In some cases, asubject may be deemed likely to have been exposed to an inflammatoryagent if the subject is diagnosed as suffering from a demyelinatingdisorder or deemed to be at increased risk for a demyelinating disorderbased on diagnostic assays and criteria described herein.

In some embodiments, one of skill in the art may use various assays,tests, diagnostic methods, or indicators of inflammation to determinewhether the compositions and methods of the present invention provideprotection of brain cells from inflammation. In some cases, indicationsof protection of cells from inflammation may comprise: reduction of thenumber or size of white matter plaques and/or lesions, improved axonalintegrity, which may be visualized through diffusion tensor imaging,reduced markers of apoptosis. In some cases, protection of cells frominflammation may comprise: reduced demyelination, reduced progressiveincrease in the number or size of white matter plaques and/or lesions,or improved remyelination. Protection of brain cells from inflammationmay also result in symptomatic improvement, e.g., improvement or delayedonset of any of the symptom(s) listed herein. In some embodiments, invitro assays may be used to determine whether the compound(s) of thepresent invention provide protection of brain cells from inflammation,for example, by administering the compound(s) to a cultured brain cellor population of cultured brain cells that have been contacted with aninflammatory agent (e.g., interferon-γ, reactive oxygen species,inflammatory cytokines), and determining viability of the cell orpopulation of cells.

In some embodiments of the present invention, the compound(s) of thepresent invention may reduce indicators of inflammation as describedherein by about 1%, about 2%, about 3%, about 4%, about 5%, about 10%,about 15%, about 20%, about 25%, about 30%, about 40%, about 50%, about60%, about 70%, about 80%, about 90%, or about 100%. In someembodiments, the compound(s) may reduce indicators of inflammation by arange of about 1-10%, about 2-20%, about 5-30%, about 10-40%, about20-100%.

Treating a Symptom(s)

In some embodiments of the present invention, the compound(s) describedherein may be used to treat a symptom of a demyelinating disorder.Exemplary symptoms of demyelinating disorders are described herein.Methods of diagnosing or monitoring symptoms of demyelinating disordersare described herein.

In some embodiments, administration of the compound(s) of the presentinvention may result in reducing the severity of symptoms by about 1%,about 2%, about 3%, about 4%, about 5%, about 10%, about 15%, about 20%,about 25%, about 30%, about 40%, about 50%, about 60%, about 70%, about80%, about 90%, or about 100%. In some embodiments, the compound(s) mayreduce the severity of symptom(s) by a range of about 1-10%, about2-20%, about 5-30%, about 10-40%, about 20-100%. In yet otherembodiments, the compound(s) may reduce the severity of symptom(s) by atleast 25%.

Delaying Onset of Symptom(s)

In some aspects, the compositions and methods of the present inventionmay be used to delay onset of symptom(s) of a demyelinating disorder.Subjects that may be a good candidate for this indication may include:subjects that have presented with one or more clinically silent lesionsas described herein, but who has not yet experienced symptom(s) of ademyelinating disorder, subjects who have experienced a single isolatedepisode but who has not yet been diagnosed with a demyelinatingdisorder, subjects who are at increased risk for developing ademyelinating disorder, or subjects that are in a remission phase of thedisorder. In some embodiments, the subject in a remission phase of ademyelinating disorder may have experienced a reduction in symptom(s)severity for at least one or at least two days, without experiencing anyworsening of any symptoms of the disorder.

In some embodiments, delayed onset of symptoms may be indicated bymonitoring the symptoms of the subject on a regular basis, e.g., bycompleting a symptom checklist about once a day. In some embodiments,delayed onset of symptoms may be indicated by physician assessment usingany of the diagnostic methods described herein. In some embodiments,delayed onset of symptoms may be indicated by comparing the frequency ofsymptom onset of the subject before and after the start of guanabenzadministration, or by comparing the frequency of symptom onset of apopulation of subjects administered guanabenz to frequency of symptomonset of a population of subjects not exposed to guanabenz and/oroptionally any other agent useful in the treatment of a demyelinatingdisorder.

In some embodiments, the methods and compositions of the presentinvention may result in a delay of symptom onset of at least about 1day, at least about 2 days, at least about 3 days, at least about 4days, at least about 5 days, at least about 6 days, at least about 1week, at least about 2 weeks, at least about 1 month, at least about 2months, at least about 4 months, at least about 6 months, at least about1 year, at least about 2 or more years.

Reducing Relapse

In some aspects, the compositions and methods of the present inventionmay be used to prevent or reduce severity of relapse of a demyelinatingdisorder. Methods for treatment regimens for reducing relapse of, andmethods for monitoring relapse of a demyelinating disorder, aredescribed herein.

In some embodiments, administration of the compound(s) described hereinmay result in a reduction of relapse incidents by about 1%, about 2%,about 4%, about 6%, about 8%, about 10%, about 20%, about 30%, about40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about100%. In some embodiments, administration of the compound(s) may reducethe frequency of relapse to an average of about 1 episode a week, about1 episode every two weeks, about 1 episode a month, about 1 episodeevery 2 months, about 1 episode every 4 months, about 1 episode every 6months, about 1 episode a year, or about less than an episode a year.

In some embodiments, the compositions and methods of the presentinvention may result in a reduction of severity of relapse. Severity ofrelapse may be indicated by monitoring the severity of symptom(s) duringrelapse. In some embodiments, administration of compound(s) describedherein may result in a reduction of relapse severity by about 1%, about2%, about 4%, about 6%, about 8%, about 10%, about 20%, about 30%, about40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%.In some embodiments, the compound(s) may reduce the severity of relapseby a range of about 1-10%, about 2-20%, about 5-30%, about 10-40%, about20-100%. In yet other embodiments, the compound(s) may reduce theseverity of relapse by at least 25%. In still yet other embodiments, thecompound(s) may reduce the severity of relapse by at least 50%.

Exemplary Compounds

The subject method utilizes a compound of Formula I:

or a derivative thereof, or a pharmaceutically acceptable salt thereof,wherein: R₁, R₂, R₃, R₄, and R₅ are independently hydrogen, deuterium,halogen, haloalkyl, alkyl, alkoxy, hydroxyl, aryl, or aryloxy. Exemplarycompounds include guanabenz and guanabenz derivatives. In someembodiments, the R1 and R5 groups are not hydrogen. In some embodiments,the R1 and R5 groups are halogen. In some embodiments, the compound isof Formula II:

In some embodiments, the compound of Formula I or its derivatives may besynthesized using methods known to those skilled in the art (see, e.g.,U.S. Pat. No. 3,975,533 or 7,932,422, herein incorporated by reference).In other embodiments, the compound of Formula II (Guanabenz, CAS No.5051-62-7) may be synthesized using methods known to those skilled inthe art (see, e.g., U.S. Pat. No. 3,982,020, Bioconjugate Chem.21:279-288, 2010, herein incorporated by reference).

Pharmaceutical Compositions

In general, the guanabenz compounds and compositions can be formulatedas pharmaceutically acceptable free base or salt forms. Pharmaceuticallyacceptable salts are described herein. In preferred embodiments, thepharmaceutically acceptable salt compositions of Formula I or Formula IIare acetate salt compositions. In other embodiments, thepharmaceutically acceptable salt compositions of Formula I or Formula IIare chlorine salt compositions.

In some embodiments, the compositions comprising an effective amount ofa compound of Formula I or Formula II may include a pharmaceuticallyacceptable carrier. In some embodiments, a pharmaceutically acceptablecarrier for the present compositions may include, but are not limitedto, amino acids, peptides, biological polymers, non-biological polymers,simple sugars or starches, inorganic salts, and gums, which may bepresent singly or in combinations thereof. In some embodiments, thepeptides used in the acceptable carrier may be gelatin. In someembodiments, the peptides used in the acceptable carrier may be albumin.In some embodiments, cellulose or its derivatives may be used in thepharmaceutically acceptable carrier. In some embodiments, the sugar usedin the acceptable carrier may be lactose. In some embodiments, the sugarused in the acceptable carrier may be glucose. In other embodiments,useful sugars may include but are not limited to, fructose, galactose,lactitol, maltitol, maltose, mannitol, melezitose, myoinositol,palatinate, raffinose, stachyose, sucrose, trehalose, xylitol, hydratesthereof, and combinations of thereof. In some embodiments, binders maybe included in the pharmaceutically acceptable carrier. Examples ofbinders include, but are not limited to, starches (for example, cornstarch or potato starch), gelatin; natural or synthetic gums such asacacia, sodium alginate, powdered tragacanth, guar gum, cellulose orcellulose derivatives (for example, methylcellulose, ethyl cellulose,cellulose acetate); microcrystalline cellulose, polyvinyl pyrrolidone,and mixtures thereof. In some embodiments, the inorganic salts used inthe acceptable carrier may be a magnesium salt, for example, magnesiumchloride or magnesium sulfate. In some embodiments, other inorganicsalts may be used, for example, calcium salts. Examples of calcium saltsinclude, but are not limited to, calcium chloride, calcium sulfate.Other examples of substances which may be used in the pharmaceuticallyacceptable carrier may include, but are not limited to, vegetable oils,such as peanut oil, cottonseed oil, olive oil, corn oil; polyols such asglycerin, propylene glycol, polyethylene glycol; pyrogen-free water,isotonic saline, phosphate buffer solutions; emulsifiers, such as theTweens®); wetting agents, lubricants, coloring agents, flavoring agents,preservatives.

The term “wetting agents” may be used interchangeably with“surfactants”, and refers to substances that lower the surface tensionof a liquid, thus allowing the liquid to spread more easily. Surfactantwhich can be used to form pharmaceutical compositions and dosage formsof the invention include, but are not limited to, hydrophilicsurfactants, lipophilic surfactants, and mixtures thereof. That is, amixture of hydrophilic surfactants may be employed, a mixture oflipophilic surfactants may be employed, or a mixture of at least onehydrophilic surfactant and at least one lipophilic surfactant may beemployed.

A suitable hydrophilic surfactant may generally have an HLB value of atleast 10, while suitable lipophilic surfactants may generally have anHLB value of or less than about 10. A useful parameter that may be usedto characterize the relative hydrophilicity and hydrophobicity ofnon-ionic amphiphilic compounds is the hydrophilic-lipophilic balance(“HLB” value). Surfactants with lower HLB values are more hydrophobic,and have greater solubility in oils, while surfactants with higher HLBvalues are more hydrophilic, and have greater solubility in aqueoussolutions. Hydrophilic surfactants are generally considered to be thosecompounds having an HLB value greater than about 10, as well as anionic,cationic, or zwitterionic compounds for which the HLB scale is notgenerally applicable. Similarly, lipophilic (i.e., hydrophobic)surfactants are generally considered to be compounds having an HLB valueequal to or less than about 10. However, HLB value of a surfactantmerely provides a rough guide generally used to enable formulation ofindustrial, pharmaceutical and cosmetic emulsions.

Hydrophilic surfactants may be either ionic or non-ionic. Suitable ionicsurfactants include, but are not limited to, alkylammonium salts, fattyacid derivatives of amino acids, glyceride derivatives of amino acids,fusidic acid salts, oligopeptides, and polypeptides, oligopeptides, andpolypeptides, lecithins and hydrogenated lecithins, lysolecithins andhydrogenated lysolecithins, phospholipids and derivatives thereof, fattyacid salts, lysophospholipids and derivatives thereof, carnitine fattyacid ester salts, salts of alkylsulfates, sodium docusate,acylactylates, mono- and di-acetylated tartaric acid esters of mono- anddi-glycerides, succinylated mono- and di-glycerides, citric acid estersof mono- and di-glycerides, and mixtures thereof.

Within the aforementioned group, ionic surfactants include, but are notlimited to, lecithins, lysolecithin, phospholipids, lysophospholipidsand derivatives thereof, carnitine fatty acid ester salts, fatty acidsalts, salts of alkylsulfates, sodium docusate, acylactylates, mono- anddi-acetylated tartaric acid esters of mono- and di-glycerides,succinylated mono- and di-glycerides, citric acid esters of mono- anddi-glycerides, and mixtures thereof.

Ionic surfactants may be the ionized forms of lactylic esters of fattyacids, lecithin, lysolecithin, phosphatidylethanolamine,phosphatidylcholine, phosphatidylglycerol, phosphatidic acid,phosphatidylserine, lysophosphatidylcholine, lysophosphatidylserine,lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidicacid, PEG-phosphatidylethanolamine, PVP-phosphatidylethanolamine,stearoyl-2-lactylate, stearoyl lactylate, succinylated monoglycerides,mono/diacetylated tartaric acid esters of mono/diglycerides, citric acidesters of mono/diglycerides, cholylsarcosine, caproate, caprylate,caprate, laurate, myristate, palmitate, oleate, linoleate, linolenate,stearate, ricinoleate, lauryl sulfate, teracecyl sulfate, docusate,lauroyl carnitines, palmitoyl carnitines, myristoyl carnitines, andsalts and mixtures thereof.

Hydrophilic non-ionic surfactants may include, but not limited to,alkylglucosides, alkylthioglucosides, alkylmaltosides, laurylmacrogolglycerides, polyoxyalkylene alkyl ethers such as polyethyleneglycol alkyl ethers, polyoxyalkylene alkylphenols such as polyethyleneglycol alkyl phenols, polyethylene glycol glycerol fatty acid esters,polyoxyalkylene alkyl phenol fatty acid esters such as polyethyleneglycol fatty acids monoesters and polyethylene glycol fatty acidsdiesters, polyglycerol fatty acid esters,polyoxyethylene-polyoxypropylene block copolymers and mixtures thereof,polyoxyalkylene sorbitan fatty acid esters such as polyethylene glycolsorbitan fatty acid esters, hydrophilic transesterification products ofa polyol with at least one member of the group consisting of glycerides,vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols,polyoxyethylene sterols and derivatives or analogues thereof,polyoxyethylated vitamins and derivatives thereof, polyethylene glycolsorbitan fatty acid esters and hydrophilic transesterification productsof a polyol with at least one member of the group consisting oftriglycerides, vegetable oils, and hydrogenated vegetable oils. Thepolyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol,propylene glycol, pentaerythritol, or a saccharide.

Other hydrophilic-non-ionic surfactants include, without limitation,PEG-10 laurate, PEG-12 laurate, PEG-12 oleate, PEG-15 oleate, PEG-20oleate, PEG-20 laurate, PEG-32 dilaurate, PEG-32 laurate, PEG-20dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20dilaurate, PEG-25 glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryllaurate, PEG-20 trioleate, PEG-30 glyceryl laurate, PEG-20 glycerylstearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30glyceryl laurate, PEG-40 glyceryl laurate, PEG-50 hydrogenated castoroil, PEG-40 castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40palm kernel oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenatedcastor oil, PEG-60 corn oil, PEG-6 caprate/caprylate glycerides, PEG-8caprate/caprylate glycerides, polyglyceryl-10 laurate, PEG-30cholesterol, PEG-25 phyto sterol, PEG-30 soya sterol, PEG-40 sorbitanoleate, PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE-9lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 oleylether, POE-20 stearyl ether, tocopheryl PEG-100 succinate, PEG-24cholesterol, polyglyceryl-10oleate, Tween 40, Tween 60, sucrosemonostearate, sucrose monolaurate, sucrose monopalmitate, PEG 10-100nonyl phenol series, PEG 15-100 octyl phenol series, and poloxamers.

Suitable lipophilic surfactants include, but are not limited to, fattyalcohols, glycerol fatty acid esters, acetylated glycerol fatty acidesters, lower alcohol fatty acids esters, propylene glycol fatty acidesters, sorbitan fatty acid esters, polyethylene glycol sorbitan fattyacid esters, sterols and sterol derivatives, polyoxyethylated sterolsand sterol derivatives, polyethylene glycol alkyl ethers, sugar ethers,sugar esters, hydrophobic transesterification products of a polyol withat least one member of the group consisting of glycerides, vegetableoils, hydrogenated vegetable oils, fatty acids and sterols, oil-solublevitamins/vitamin derivatives, lactic acid derivatives of mono- anddi-glycerides, and mixtures thereof. Within this group, preferredlipophilic surfactants include glycerol fatty acid esters, propyleneglycol fatty acid esters, and mixtures thereof, or are hydrophobictransesterification products of a polyol with at least one member of thegroup consisting of vegetable oils, hydrogenated vegetable oils, andtriglycerides.

In some embodiments, lubricants that may be used in the pharmaceuticalcomposition include, but are not limited to, agar, calcium stearate,magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol,mannitol, polyethylene glycol, other glycols, stearic acid, sodiumlauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil,cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethylaureate, or mixturesthereof. Additional lubricants include, by way of example, a syloidsilica gel, a coagulated aerosol of synthetic silica, or mixturesthereof. A lubricant can optionally be added, in an amount of less thanabout 1 weight percent of the pharmaceutical composition.

In some embodiments, the composition may include a solubilizer to ensuregood solubilization of the compound and to reduce precipitation of thecompound of the present invention. A solubilizer may be used to increasesolubility of the compound or other active ingredients, or may be usedto maintain the composition as a homogeneous solution or dispersion.Examples of suitable solubilizers include but are not limited to,alcohols and polyols such as ethanol, isopopropanol, polyvinyl alcohol,gelatin, mannitol, sodium carboxymethyl cellulose (CMCNa), povidone,propylene glycol, polyethylene glycol, polyvinyl pyrolidone, glycerin,cyclodextrins or cyclodextrin derivatives, polyethylene glycol ethers ofmolecular weight averaging about 200 to about 6000, such as PEG, amidesand other nitrogen-containing compounds such as 2-pyrrolidone,2-piperidone, epsilon.-caprolactam, N-alkylpyrrolidone,N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam,dimethylacetamide and polyvinylpyrrolidone, esters such as ethylpropionate, tributylcitrate, acetyl triethylcitrate, acetyl tributylcitrate, triethylcitrate, ethyl oleate, ethyl caprylate, ethyl butyrate,triacetin, propylene glycol monoacetate, propylene glycol diacetate,ε-caprolactone and isomers thereof, δ-valerolactone and isomers thereof,β-butyrolactone and isomers thereof, and other solubilizers known in theart, such as dimethyl acetamide, dimethyl isosorbide, N-methylpyrrolidones, monooctanoin, diethylene glycol monoethyl ether, water, ormixtures and/or combinations thereof.

Mixtures of solubilizers may also be used. Examples include, but notlimited to, ethyl oleate, ethyl caprylate, triacetin, triethylcitrate,dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone,polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropylcyclodextrins, ethanol, polyethylene glycol 200-100, transcutol,propylene glycol, glycofurol and dimethyl isosorbide. Particularlypreferred solubilizers include sorbitol, glycerol, triacetin, ethylalcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that can be included is not particularlylimited. The amount of a given solubilizer may be limited to abioacceptable amount, which may be readily determined by one of skill inthe art. In some circumstances, it may be advantageous to includeamounts of solubilizers far in excess of bioacceptable amounts, forexample, to maximize the concentration of the drug, with excesssolubilizer removed prior to providing the composition to a subjectusing conventional techniques, such as distillation or evaporation.Thus, if present, the solubilizer can be in a weight ratio of 10%, 25%,50%, 75%, 100%, or up to about 200% by weight, based on the combinedweight of the drug, and other excipients. If desired, very small amountsof solubilizer may also be used, such as 5%, 2%, 1%, 0.5% or even less.Typically, the solubilizer may be present in an amount of about 1% toabout 100%, more typically about 5% to about 25% by weight.

In some embodiments, the composition may include one or morepharmaceutically acceptable additives, which may include, but are notlimited to, detackifiers, anti-foaming agents, buffering agents,antioxidants, polymers, preservatives, chelating agents, odorants,opacifiers, suspending agents, fillers, plasticizers, and mixturesthereof.

In some embodiments, the pharmaceutically acceptable carrier comprisesmore than 90%, more than 80%, more than 70%, more than 60%, more than50%, more than 40%, more than 30%, more than 20%, more than 10%, morethan 9%, more than 8%, more than 6%, more than 5%, more than 4%, morethan 3%, more than 2%, more than 1%, more than 0.5%, more than 0.4%,more than 0.3%, more than 0.2%, more than 0.1%, more than 0.09%, morethan 0.08%, more than 0.07%, more than 0.06%, more than 0.05%, more than0.04%, more than 0.03%, more than 0.02%, more than 0.01%, more than0.009%, more than 0.008%, more than 0.007%, more than 0.006%, more than0.005%, more than 0.004%, more than 0.003%, more than 0.002%, more than0.001%, more than 0.0009%, more than 0.0008%, more than 0.0007%, morethan 0.0006%, more than 0.0005%, more than 0.0004%, more than 0.0003%,more than 0.0002%, or more than 0.0001% of the pharmaceuticalcomposition by w/w, w/v or v/v.

In some embodiments, the concentration of the compound of Formula I orFormula II in the composition comprises less than 100%, less than 90%,less than 80%, less than 70%, less than 60%, less than 50%, less than40%, less than 30%, less than 20%, less than 10%, less than 9%, lessthan 8%, less than 6%, less than 5%, less than 4%, less than 3%, lessthan 2%, less than 1%, less than 0.5%, less than 0.4%, less than 0.3%,less than 0.2%, less than 0.1%, less than 0.09%, less than 0.08%, lessthan 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than0.03%, less than 0.02%, less than 0.01%, less than 0.009%, less than0.008%, less than 0.007%, less than 0.006%, less than 0.005%, less than0.004%, less than 0.003%, less than 0.002%, less than 0.001%, less than0.0009%, less than 0.0008%, less than 0.0007%, less than 0.0006%, lessthan 0.0005%, less than 0.0004%, less than 0.0003%, less than 0.0002%,or less than 0.0001% of the pharmaceutical composition by w/w, w/v orv/v.

In some embodiments, the concentration of the compound of Formula I orFormula II is in the range of about 0.0001% to about 50%, about 0.001%to about 40%, about 0.01% to about 20%, about 0.02% to about 29%, about0.03% to about 28%, about 0.04% to about 27%, about 0.05% to about 26%,about 0.06% to about 25%, about 0.07% to about 24%, about 0.08% to about23%, about 0.09% to about 22%, about 0.1% to about 21%, about 0.2% toabout 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5%to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about0.8% to about 14%, about 0.9% to about 12%, about 1% to about 10% of thepharmaceutical composition by w/w, w/v or v/v. v/v.

In some embodiments, the concentration of the compound of Formula I orFormula II is in the range of about 0.0001% to about 5%, about 0.001% toabout 4%, about 0.01% to about 2%, about 0.02% to about 1%, or about0.05% to about 0.5% of the pharmaceutical composition by w/w, w/v orv/v. v/v.

In some embodiments, the amount of the compound of Formula I or FormulaII in the pharmaceutical composition is about 0.00001 mg, 0.0001 mg,0.001 mg, 0.005 mg, 0.01 mg, 0.05 mg, 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2mg, 4 mg, about 8 mg, about 16 mg, about 32 mg, about 64 mg, about 100mg, about 200 mg, about 500 mg, about 1 g, about 2 g, about 5 g, about10 g.

In some embodiments, the pharmaceutical compositions of Formula I orFormula II may be manufactured by Ivax Pharmaceuticals. In someembodiments, the pharmaceutical compositions of Formula I or Formula IImay be manufactured by Teva Pharmaceuticals. In some embodiments, thepharmaceutical compositions of Formula I or Formula II may bemanufactured by Sandoz Inc. In some embodiments, the pharmaceuticalcompositions of Formula I or Formula II may be manufactured by WatsonLaboratories Inc. In some embodiments, the pharmaceutical compositionsof Formula I or Formula II may be manufactured by Wyeth AyerstLaboratories.

In some cases, the pharmaceutical compositions of Formula I or FormulaII may be packaged by Ivax Pharmaceuticals. In some cases, thepharmaceutical compositions of Formula I or Formula II may be packagedby Letco Medical Inc. In some cases, the pharmaceutical compositions ofFormula I or Formula II may be packaged by Murfreesboro Pharmaceuticals.In some cases, pharmaceutical compositions of Formula I or Formula IImay be packaged by the Pharmaceutical Utilization Management ProgramVA., Inc.

In some cases, the pharmaceutical compositions of Formula I or FormulaII may comprise an additional agent. The additional agent may be anagent useful in the treatment of a demyelinating disorder. Examples ofother agents useful in the treatment of demyelinating disorders include,but are not limited to, interferon β, IFN β-1a (brand names: AVONEX,REBIF), IFNβ-1b (brand name: BETASERON), glatiramer acetate (COPAXONE,Copolymer-1), natalizumab (TYSABRI), mitoxantrone (C₂₂H₂₈N₄O₆.2HCl),methylprednisone, methylprednisolone, thalidomide, fingolimod (GILENIA),dimethyl fumarate (BG-12), teriflunomide (AUBAGIO), anti-LINGO antibody,alemtuzumab (CAMPATH), dalfampridine, PEG-interferon beta-1a (BIIB017),daclizumab (ZENAPAX), laquinimod, and ocrelizumab. Accordingly, apharmaceutical composition of Formula I or Formula II may comprise oneor more agents selected from the group consisting of interferon β, IFNβ-1a (brand names: AVONEX, REBIF), IFNβ-1b (brand name: BETASERON),glatiramer acetate (COPAXONE, Copolymer-1), natalizumab (TYSABRI),mitoxantrone (C₂₂H₂₈N₄O₆.2HCl), methylprednisone, methylprednisolone,thalidomide, fingolimod (GILENIA), dimethyl fumarate (BG-12),teriflunomide (AUBAGIO), anti-LINGO antibody, alemtuzumab (CAMPATH),dalfampridine, PEG-interferon beta-1a (BIIB017), daclizumab (ZENAPAX),laquinimod, and ocrelizumab.

Described below are some non-limiting examples of pharmaceuticalcompositions.

Pharmaceutical Compositions for Oral Administration

In some embodiments, the invention provides a pharmaceutical compositioncomprising an effective amount of a compound of Formula I or Formula IIfor oral administration containing at least one therapeutic agent and apharmaceutically acceptable carrier for oral administration. In someembodiments, pharmaceutically acceptable carriers described herein canbe suitable for oral administration.

In some embodiments, the pharmaceutical composition comprising aneffective amount of a compound of Formula I or Formula II for oraladministration is a solid pharmaceutical composition. In someembodiments, the solid pharmaceutical composition may be presented asdiscrete oral dosage forms. Non-limiting examples of discrete oraldosage forms include tablets, capsules, caplets, gelatin capsules,sustained release formulations, lozenges, thin films, lollipops, chewinggum.

In some embodiments, discrete oral dosage forms such as tablets may becoated by known techniques to delay or prolong absorption in thegastrointestinal tract, thus providing a sustained action of a longerperiod of time. In some cases, such sustained release may be beneficialto reduce side-effects, such as drowsiness. In some embodiments, thecompound of Formula I or Formula II may be mixed with one or more inertsolid diluents, such as calcium carbonate or calcium phosphate. In someembodiments, the compound of Formula I or Formula II may be presented assoft gelatin capsules, wherein the compound is mixed with water or anoil medium, such as peanut oil, or olive oil, for example.

In some embodiments, the pharmaceutical composition comprising aneffective amount of a compound of Formula I or Formula II for oraladministration is a liquid pharmaceutical composition. Non-limitingexamples of liquid compositions for oral administration includehydrophilic suspensions, emulsions, liquids, gels, syrups, slurries,solutions, elixirs, softgels, tinctures, hydrogels.

In some embodiments, solid or liquid compositions comprising aneffective amount of a compound of Formula I or Formula II for oraladministration may also comprise various sweetening or flavoring agents,or coloring agents. Examples of coloring agents include dyes suitablefor food such as those known as F.D. & C. dyes and natural coloringagents such as grape skin extract, beet red powder, beta carotene,annato, carmine, turmeric, paprika, and so forth. Derivatives,analogues, and isomers of any of the above colored compound also may beused.

Such dosage forms may be prepared by methods well known to those skilledin the art, e.g., in a pharmacy. Such methods would comprise bringingthe compound of Formula I or Formula II into association with thepharmaceutically acceptable carrier.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising an effective amount of a compound of FormulaI or Formula II, since water may facilitate the degradation of thecompounds. In some embodiments, the anhydrous pharmaceuticalcompositions and dosage forms of the invention may be prepared usinganhydrous or low moisture containing ingredients. In some embodiments,the anhydrous pharmaceutical compositions and dosage forms of theinvention may be prepared under low humidity or low moisture conditions.In some embodiments, the pharmaceutical compositions of the presentinvention which contain lactose may be made anhydrous if substantialcontact with moisture and/or humidity during manufacturing, packaging,and/or storage is expected. In some embodiments, an anhydrouspharmaceutical composition comprising an effective amount of a compoundof Formula I or Formula II may be prepared and stored such that itsanhydrous nature is maintained. For example, the anhydrous compositionsmay be packaged using materials known to prevent exposure to water suchthat they can be included in suitable formulary kits, examples of whichinclude, but are not limited to, hermetically sealed foils, plastic orthe like, unit dose containers, blister packs, and strip packs.

Pharmaceutical Compositions for Injection or Parenteral Administration

In some aspects, the present invention provides pharmaceuticalcompositions comprising an effective amount of a compound of Formula Ior Formula II for parenteral administration for the treatment of variousdemyelinating disorders. “Parenteral administration” refers to routes ofadministration other than the gastro-intestinal tract. Examples ofparenteral administration include, but are not limited to, intravenousinjection, subcutaneous injection, intramuscular injection, infusion, orimplantation. In some embodiments, infusion may be intradermal, orsubcutaneous, or through a transdermal implant. In some cases, infusionmay be intracerebral, intracerebrovascular, or epidural. Pharmaceuticalcompositions for parenteral administration are well known in the art.Examples of the compositions for parenteral administration are disclosedin the following references which are hereby incorporated by reference:US 2006/0287221, U.S. Pat. Nos. 5,244,925, 4,309,421, 4,158,707,5,164,405).

In some embodiments, compositions comprising an effective amount of acompound of Formula I or Formula II for parenteral administration mayinclude aqueous solutions and/or buffers commonly used for injectionand/or infusion. Commonly used aqueous buffers and/or solutions mayinclude, but are not limited to sodium chloride solutions of about 0.9%,phosphate buffers, Lactated Ringer's solution, Acetated ringer'ssolution, phosphate buffered saline, citrate buffers, Tris buffers,histidine buffers, HEPES buffers, glycine buffers, N-glycylglycinebuffers, and the like. Other pharmaceutically acceptable carriers forparenteral administration may include ethanol, glycerol, propyleneglycol, cyclodextrin and cyclodextrin derivatives, vegetable oils, andthe like.

In some embodiments, pharmaceutical compositions comprising an effectiveamount of a compound of Formula I or Formula II for injection and/orinfusion may also contain preservatives present in amounts thateffectively prevent or reduce microbial contamination or degradation.Various agents, e.g., phenol, m-cresol, benzyl alcohol, parabens,chlorobutanol, methotrexate, sorbic acid, thimerosol, ethylhydroxybenzoate, bismuth tribromophenate, methyl hydroxybenzoate,bacitracin, propyl hydroxybenzoate, erythromycin, 5-fluorouracil,doxorubicin, mitoxantrone, rifamycin, chlorocresol, benzalkoniumchlorides, may be used to prevent or reduce contamination.

In some embodiments, sterile solutions may be prepared by incorporatingthe compound of Formula I and/or II in the required amount in theappropriate solvent with various other ingredients as described herein,as required, followed by filtered sterilization. Generally, dispersionsare prepared by incorporating the various sterilized active ingredientsinto a sterile vehicle which contains the basic dispersion medium andthe required other ingredients from those enumerated above. In the caseof sterile powders for the preparation of sterile injectable solutions,certain methods of preparation include but are not limited tovacuum-drying and freeze-drying techniques which yield a powder of theactive ingredient plus any additional desired ingredient from apreviously sterile-filtered solution thereof.

Pharmaceutical Compositions for Transdermal (Topical) Delivery

In some embodiments, the invention provides a pharmaceutical compositioncomprising an effective amount of a compound of Formula I or Formula IIfor transdermal delivery, and a pharmaceutical excipient suitable fortransdermal delivery. Compositions of the present invention can beformulated into preparations in liquid, semi-solid, or solid formssuitable for local or topical administration. Examples of forms suitablefor topical or local administration include but are not limited to,gels, water soluble jellies, creams, lotions, suspensions, foams,powders, slurries, ointments, oils, pastes, suppositories, solutions,sprays, emulsions, saline solutions, dimethylsulfoxide (DMSO)-basedsolutions. In general, carriers with higher densities are capable ofproviding an area with a prolonged exposure to the active ingredients.In contrast, a solution formulation may provide more immediate exposureof the active ingredient to the chosen area.

The pharmaceutical compositions comprising an effective amount of acompound of Formula I or Formula II also may comprise suitable solid orgel phase carriers, which are compounds that allow increased penetrationof, or assist in the delivery of, therapeutic molecules across thestratum corneum barrier of the skin. There are many of thesepenetration-enhancing molecules known to those skilled in the art oftopical formulation. Examples of such carriers and excipients include,but are not limited to, alcohols (e.g., ethanol), fatty acids (e.g.,oleic acid), humectants (e.g., urea), glycols (e.g., propylene glycol),surfactants (e.g., isopropyl myristate and sodium lauryl sulfate),glycerol monolaurate, sulfoxides, pyrrolidones, terpenes (e.g.,menthol), amines, amides, alkanes, alkanols, water, calcium carbonate,calcium phosphate, various sugars, starches, cellulose derivatives,gelatin, and polymers such as polyethylene glycols.

Another exemplary formulation for use in the methods of the presentinvention employs transdermal delivery devices (“patches”). Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of the compounds of the present invention in controlledamounts, either with or without another agent. The construction and useof transdermal patches for the delivery of pharmaceutical agents is wellknown in the art. See, e.g., U.S. Pat. Nos. 5,023,252, 4,992,445 and5,001,139, which are herein incorporated by reference.

Pharmaceutical Compositions for Inhalation.

In some embodiments, the invention provides a pharmaceutical compositioncomprising an effective amount of a compound of Formula I or Formula IIfor transdermal delivery, and a pharmaceutical excipient suitable fordelivery by inhalation. Compositions for inhalation include solutionsand suspensions in pharmaceutically acceptable, aqueous or organicsolvents, or mixtures thereof, and powders. The liquid or solidcompositions may contain suitable pharmaceutically acceptable excipientsas described herein. The compositions may be administered by the oral ornasal respiratory route for systemic effect. In some embodiments,compositions in preferably pharmaceutically acceptable solvents may benebulized by use of inert gases. In some embodiments, nebulizedsolutions may be inhaled directly from the nebulizing device. In otherembodiments, nebulizing device may be attached to a face mask tent orintermittent positive pressure breathing machine. Solution, suspension,or powder compositions may be administered, preferably orally ornasally, from devices that deliver the formulation in an appropriatemanner.

Other Pharmaceutical Compositions.

Pharmaceutical compositions comprising the compound of Formula I and/orII may also be prepared from compositions described herein and one ormore pharmaceutically acceptable excipients suitable for buccal,sublingual, rectal, intraosseous, intraocular, intranasal, epidural, orintraspinal administration. Preparations for such pharmaceuticalcompositions are well-known in the art. See, e.g., Anderson, Philip O.;Knoben, James E.; Troutman, William G, eds., Handbook of Clinical DrugData, Tenth Edition, McGraw-Hill, 2002; Pratt and Taylor, eds.,Principles of Drug Action, Third Edition, Churchill Livingston, N.Y.,1990; Katzung, ed., Basic and Clinical Pharmacology, Ninth Edition,McGraw Hill, 20037ybg; Goodman and Gilman, eds., The PharmacologicalBasis of Therapeutics, Tenth Edition, McGraw Hill, 2001; RemingtonsPharmaceutical Sciences, 20th Ed., Lippincott Williams & Wilkins., 2000;Martindale, The Extra Pharmacopoeia, Thirty-Second Edition (ThePharmaceutical Press, London, 1999); all of which are incorporated byreference herein in their entirety.

Exemplary Treatment Regimens and Routes of Administration

Administration of each compounds or pharmaceutical composition of thepresent invention can be effected by any method that enables delivery ofthe compounds to the site of action. In some embodiments, the route ofadministration may be oral, by intraperitoneal injection, by inhalation,by transdermal infusion, or parenteral. The preferred route ofadministration is oral. In some cases, the oral administration maycomprise administration of any of the oral dosage forms as describedherein. The effective amount of the compound of Formula I or Formula IIadministered will be dependent on the subject being treated, theseverity of the disorder or condition, the subject's risk of developinga demyelinating disorder, the rate of administration, the disposition ofthe compound and the discretion of the prescribing physician. However,an effective dosage is in the range of about 0.001 to about 1 mg per kgbody weight per day, preferably about 0.01 to about 0.5 mg/kg/day, insingle or divided doses. For a 70 kg human, this would amount to about0.1 to about 64 mg/day, or about 2 to about 32 mg/day. In someinstances, dosage levels below the lower limit of the aforesaid rangemay be more than adequate, while in other cases still larger doses maybe employed without causing any harmful side effect, e.g., by dividingsuch larger doses into several small doses for administration throughoutthe day.

In some cases, as for administration to humans, the oral dosage form maycomprise 0.1, 0.5, 1, 2, 4, 8, 16, 24, 32, or 64 mg of the compound ofFormula I and/or II. In preferred cases, the oral dosage form is atablet. In some cases, the oral administration may compriseadministration of a liquid formula.

In some embodiments, the route of administration is by injection. Insome embodiments, the injection is intraperitoneal. In some embodiments,the route of administration is intravenous. In some embodiments, theinjection is subcutaneous. In some embodiments, the injection isintramuscular. In some cases, as for administration to humans, theinjection or injected dosage form may comprise 0.1, 0.5, 1, 2, 4, 8, 16,24, 32, or 64 mg of the compound of Formula I and/or II.

In some embodiments, administration may comprise inhalation. In somecases, as for administration to humans, the inhaled dose or dosage formmay comprise 0.1, 0.5, 1, 2, 4, 8, 16, 24, 32, or 64 mg of the compoundof Formula I and/or II.

In some embodiments, administration may comprise infusion. In somecases, infusion may involve chronic, steady dosing. Devices for chronic,steady dosing, i.e.—by a controlled pump, are known in the art,(examples may be described in U.S. Pat. Nos. 7,341,577, 7,351,239,8,058,251, herein incorporated by reference). In some cases,administration may comprise intracerebrovascular delivery. In somecases, administration may comprise intracerebral delivery. Methods forintracerebrovascular or intracerebral delivery are well known in theart, as disclosed by the following references, herein incorporated byreference: U.S. Pat. Nos. 5,711,316, 5,713,923, 5,735,814, 5,832,702,5,836,935, 5,720,720, Chandler et al., Ann. N.Y. Acad. Sci., 531, 1988,pp. 206-212, Bouvier et al, Neurosurgery 20(2), 1987, pp. 286-291,Johnson et al., Ann. N.Y. Acad. Sci., 531, 1988, pp. 57-67, andSendelbeck et al., Brain Res., 328, 1985, pp. 251-258, hereinincorporated by reference. In some cases, administration may comprisedirect injection into the spinal cord, for example, by epiduralinjection.

In some embodiments, one oral or injected dose for humans may be about0.1 mg, about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg,about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 10 mg, about 12mg, about 16 mg, about 32 mg, or about 64 mg. In some embodiments, oneoral or injected dose for non-human subjects may be about 0.001 mg/kg,about 0.005 mg/kg, about 0.01 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg,about 0.5 mg/kg, about 1 mg/kg, about 2 mg/kg, about 4 mg/kg, about 8mg/kg, about 16 mg/kg, about 32 mg/kg, about 64 mg/kg. In some cases, anoral or injected dose may be administered about once per day, abouttwice per day, about 3-5 times per day, about once every other day,about once every two days, about twice a week, about once a week, aboutonce every two weeks, about once a month, for as long as the subject isin need thereof.

One dose administered through intracerebrovascular delivery orintracerebral delivery may be about 50 ng, about 100 ng, about 200 ng,about 500 ng, about 1 μg, about 10 μg, about 25 μg, about 50 μg, about100 μg, about 200 μg, about 500 μg, about 1 mg, about 5 mg, about 10 mg.In some cases, a dose may be administered about once per day, about onceevery other day, about twice a week, about once a week, about once everytwo weeks, about once a month, about once every 2-6 months, for as longas the subject is in need thereof.

In some cases, administration of guanabenz or its derivatives maycommence for a subject in need thereof when the subject is firstsuspected of having a demyelinating disorder. In some cases,administration of guanabenz or its derivatives may commence for asubject in need thereof when the subject diagnosed with a demyelinatingdisorder. In some cases, administration of guanabenz or its derivativesmay commence for a subject in need thereof before the onset of anysymptoms of a demyelinating disorder, or after symptom onset. In somecases, administration of guanabenz or its derivatives may commence for asubject in need thereof after the subject has experienced a singleisolated clinical symptom or symptoms. In some cases, administration ofguanabenz or its derivatives may commence for a subject in need thereofwhen the subject is deemed to be at increased risk for developing ademyelinating disorder. In some cases, administration of guanabenz orone of its derivatives may commence for a subject in need thereof whenthe subject is in a remission phase of the disorder, for the purpose ofpreventing or reducing the severity of relapse. In some cases, a subjectin remission may be administered guanabenz or one of its derivativesafter symptom severity has reached a peak and has been in decline for atleast one or two or several days.

Administration of the compound(s) of the invention may continue as longas necessary. In some embodiments, the compound(s) invention isadministered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In someembodiments, the compound(s) invention is administered for more than 1month, more than 2 months, more than 4 months, more than 6 months, morethan 1 year, more than 2 years, more than 5 years. In some embodiments,the compound(s) of the invention is administered for less than for lessthan 1 month, less than 2 months, less than 4 months, less than 6months, less than 1 year, less than 2 years, less than 5 years. In someembodiments, the compound(s) of the invention is administered for lessthan 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, an agentof the invention is administered chronically on an ongoing basis, e.g.,for the treatment or prevention of a demyelinating disorder.

The compounds of the invention may be administered in dosages. It isknown in the art that due to inter-subject variability in compoundpharmacokinetics, individualization and/or adjustment of the dosingregimen may be necessary for optimal therapy.

In some cases, the timing of dosing may be chosen in order to minimizepotential side-effects from the compounds of the invention. For example,the compounds of the invention may, in some instances, induce drowsinessin a subject. In some embodiments, a dose of a compound of the inventionis provided in the evening, in order to minimize the effect ofdrowsiness on the activities of the subject which, in turn, may allow ahigher dose to be administered to the subject in a single dosage. Insome embodiments, a single dose of about 64 mg of a compound of theinvention is administered in the evening. As used herein, the term“evening” generally refers to a time after about 5:00 pm, after about6:00 pm, after about 7:00 pm, after about 8:00 pm, after about 9:00 pm,after about 10:00 pm, or after about 11:00 pm. In some embodiments, acompound of the invention may be administered with another compound thatcounteracts drowsiness, such as a stimulant.

Exemplary Combination Therapies

In some aspects of the present invention, the compounds described hereincan also be used in combination with other known agents that areselected for their therapeutic value to the subject in need thereof. Insome aspects of the present invention, the compound(s) described hereinmay be used in a combination therapy which comprises one or moreadditional agents known to modulate other pathways, or other componentsof the same pathways as that modulated by the compound(s), or the sametargets of the same pathway modulated by the compound(s). Examples ofpathways or pathway components modulated by the compound(s) describedherein and may also be targeted by the one or more additional agentsinclude, but are not necessarily limited to, ER stress pathways, PERKpathways, eIF2α, phosphorylated eIF2α, GADD34, protein phosphatase I(PPI), ATF4, IRE1 pathways, ATF6 pathways. Examples of other pathways orpathway components that may be targeted by the one or more additionalagents include, but are not limited to: pathways related to immunefunction, immunomodulatory pathways, inflammatory pathways, cytokinepathways, T cells, T cell proliferation pathways, CD4+ T cells, Tregcells, CD8+ T cells, Major Histocompatibility Complex (MHC), antigenpresenting cells (APC), adhesion molecule, MMP and/or chemokineexpression, leukocytes, leukocyte migration across the BBB, α4integrins, lymphocytes, monocytes, topisomerase-2, macrophages, B cells,B or T lymphocytes, interleukins, TNF-α, interferons (IFN), IFN-β,IFN-γ, neurotrophic pathways, neurotrophic factors such as, e.g., NGF,BDNF, NT-3, CNTF, neuroprotective pathways, immune tolerance to myelinor myelin-associated proteins, antibody production, etc., apoptosis,protein folding, chaperones, heat shock proteins, etc. Of particularinterest are agents known to be useful for the treatment ofdemyelinating disorders.

In some embodiments of the use of the compounds described herein forcombination therapy, other agents may be administered in the samepharmaceutical composition. In some embodiments of the use of thecompounds described herein for combination therapy, other agents do nothave to be administered in the same pharmaceutical composition, but maybe administered simultaneously with the composition of the compound(s)described herein. In other embodiments of the use of the compoundsdescribed herein for combination therapy, the other agents may beadministered at different times. In some embodiments, because ofdifferences in physical and chemical characteristics of the compound(s)described herein and the other agents, the compound(s) and other agentsmay be administered by different routes. The determination of the modeof administration and the advisability of administration in the samepharmaceutical composition is well within the knowledge of one skilledin the art, e.g., a physician. The initial co-administration can be madeaccording to established protocols known in the art, and then, basedupon the observed effects, the dosage, modes of administration and timesof administration can be modified by the skilled clinician.

In some embodiments, it may be appropriate to administer at least one ofthe compounds described herein in combination with another therapeuticagent. In a non-limiting example, if a subject is experiencing a sideeffect from administration of one of the compounds described herein(such as, for example, drowsiness), then it may be appropriate toadminister an anti-drowsiness agent in combination with the compound.Alternatively, by way of another non-limiting example, the therapeuticeffectiveness of one of the compounds described herein may be enhancedby administration of an adjuvant (wherein the adjuvant may have minimaltherapeutic benefit, but in combination with another therapeutic agentsuch as the compound(s) described herein, the overall therapeuticbenefit to the patient is enhanced). In another non-limiting example,the therapeutic benefit to a subject may be increased or enhanced by acombination therapy of one of the compounds described herein with one ormore additional therapeutic agents that also has therapeutic benefit. Insome embodiments, the overall benefit experienced by the patient fromthe combination therapy may be an additive effect of the individualagents. In some embodiments, the subject may experience a synergisticbenefit from the combination therapy.

One of skill in the art will appreciate that therapeutically effectivedosages may vary when the agents are used in combination therapy.Methods for determining therapeutically effective dosages of drugs andother agents for use in combination therapy regimens are known to thoseof skill in the art.

It is understood by those of skill in the art that the combinationtherapy regimen for the subject in need thereof can be modified inaccordance with a variety of factors. These factors may include, but arenot limited to, the demyelinating disorder from which the subjectsuffers from or is at increased risk for, any other medical conditionexperienced by the subject, as well as the age, weight, sex, diet, andmedical history of the subject. Thus, the combination therapy regimenactually employed can vary widely and therefore can deviate fromexemplary combination therapies described herein.

Additional agents useful for combination therapy include any agentscapable of modulating a target molecule implicated in a demyelinatingdisorder, either directly or indirectly. Non-limiting examples of targetmolecules modulated by additional agents include enzymes, enzymesubstrates, antibodies, antigens, membrane proteins, products oftransitions, nuclear proteins, lipid rafts, cytosolic proteins,mitochondrial proteins, lysosomal proteins, scaffold proteins,phosphoproteins, glycoproteins, nuclear receptors, membrane receptors,G-protein-coupled receptors, nuclear receptors, protein tyrosinekinases, protein serine/threonine kinases, phosphatases, proteases,hydrolases, lipases, phospholipases, ligases, reductases, oxidases,synthases, transcription factors, ion channels, RNA, DNA, RNAse, DNAse,phospholipids, sphingolipids, ion channel proteins, nucleotide-bindingproteins, calcium-binding proteins, chaperones, DNA binding proteins,RNA binding proteins, scaffold proteins, tumor suppressors, cell cycleproteins, and histones.

Additional agents may be: small molecules, nutraceuticals, vitamins,e.g., vitamin D, drugs, pro-drugs, biologics, peptides, peptidemimetics, antibodies, antibody fragments, cell or tissue transplants,vaccines, polynucleotides, DNA molecules, RNA molecules, (i.e.—siRNA,miRNA), antibodies conjugated to drugs, toxins, fusion proteins. Agentsmay be delivered by vectors, including but not limited to: plasmidvectors, viral vectors, non-viral vectors, liposomal formulations,nanoparticle formulations, toxins, therapeutic radioisotopes, etc.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with one or more agents that are currentlyapproved by the FDA for the treatment or prophylaxis of a demyelinatingdisorder. Examples of other agents approved by the FDA for treatment orprophylaxis of demyelinating disorders include, but are not limited to,interferon β, IFN β-1a (brand names: AVONEX, REBIF), IFNβ-1b (brandname: BETASERON), glatiramer acetate (COPAXONE, Copolymer-1),natalizumab (TYSABRI), mitoxantrone (C₂₂H₂₈N₄O₆.2HCl), methylprednisone,methylprednisolone, thalidomide, fingolimod (GILENIA), dimethyl fumarate(BG-12), teriflunomide (AUBAGIO), anti-LINGO antibody, alemtuzumab(CAMPATH), dalfampridine, PEG-interferon beta-1a (BIIB017), daclizumab(ZENAPAX), laquinimod, and ocrelizumab. Accordingly, a method of theinvention may comprise administering to a subject a compound of thepresent invention with one or more additional therapeutic agent usefulin the treatment and/or prophylaxis of a demyelinating disorder.Compound(s) of the present invention may be administered before, after,or concurrently with the one or more additional therapeutic agents. Insome embodiments, compound(s) of the present invention may be formulatedin a composition comprising a pharmaceutically acceptable carrier andthe one or more additional therapeutic agents. In one embodiment, thecompound(s) of the present invention are administered in combinationwith glatiramer acetate (COPAXONE). In one embodiment, the compound(s)of the present invention are administered in combination with dimethylfumerate. In one embodiment, the compound(s) of the present inventionare administered in combination with fingolimod. In one embodiment, thecompound(s) of the present invention are administered in combinationwith interferon beta-1a. In one embodiment, the compound(s) of thepresent invention are administered in combination with interferonbeta-1b. In one embodiment, the compound(s) of the present invention areadministered in combination with mitoxantrone. In one embodiment, thecompound(s) of the present invention are administered in combinationwith natalizumab. In one embodiment, the compound(s) of the presentinvention are administered in combination with dalfampridine. In oneembodiment, the compound(s) of the present invention are administered incombination with daclizumab. In some embodiments the compound isguanabenz. In some embodiments, the formulation may be a formulation forinjection. In some embodiments the formulation may be a formulation forsubcutaneous administration. In some embodiments the formulation may bea formulation for intravenous administration.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are associated withaltered clinical outcomes of a demyelinating disorder but not FDAapproved for the treatment of the disorder. Examples of other agentsassociated with altered clinical outcomes of a demyelinating disorderbut not approved for treatment of the disorder include, but are notlimited to, cyclophosphamide, methotrexate, azathioprine, cyclosporine,etc.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are associated withneuroprotection or axonal regeneration. Examples of such agents include,but are not limited to, neurotrophic factors (e.g., NT-3, BDNF, NGF,CTNF, FGF, GDNF), MANF polypeptides, stem cell transplantation therapy(e.g., transplantation of hematopoietic and non-hematopoietic stemcells, agents that target pathways associated with neurodegeneration(e.g., ubiquitin-proteasome pathways, ER stress pathways, autophagypathways, mitochondrial pathways, oxidative stress pathways, apoptosispathways, etc.). Further examples of pathways, pathway components, oragents that may promote axonal regeneration include, but are not limitedto, PirB pathways, Nogo receptor pathways, netrin, bone morphogenicprotein/Smad1 pathways, cAMP, Vitamin D, among others.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are useful forreducing or preventing damage induced by inflammation. Examples of suchagents include but are not necessarily limited to; steroidalanti-inflammatory drugs, non-steroidal anti-inflammatory drugs, immuneselective anti-inflammatory drugs, cyclooxygenase (COX) inhibitors,prostaglandin inhibitors, herbs with anti-inflammatory qualities (e.g.,Harpagophytum, hyssop, ginger, turmeric, Arnica montana, sesquiterpenelactone, willow bark), among others.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are useful for thetreatment of pain and/or inflammation symptoms that may be associatedwith a demyelinating disorder. Examples of such agents include, but arenot limited to, analgesics, histamine, histamine antagonists,bradykinin, bradykinin antagonists, agents that modulate5-hydroxytryptamine (serotonin) pathways, serotonin reuptake inhibitors,serotonin-norepinephrine reuptake inhibitors, tricyclic antidepressants,gamma-aminobutyric acid analogs, gamma-aminobutyric acid receptoragonists, benzodiazapenes, narcotics, eicosanoids, prostaglandins,thromboxanes, leukotrienes, aspirin, anti-inflammatory agents,nonsteroidal anti-inflammatory agents, analgesic-antipyretic agents,agents that inhibit the synthesis of prostaglandins and thromboxanes,selective inhibitors of the inducible cyclooxygenase, selectiveinhibitors of the inducible cyclooxygenase-2, autacoids, paracrinehormones, somatostatin, gastrin, agents that target cytokines whichmediate interactions involved in humoral and cellular immune responses,lipid-derived autacoids, β-adrenergic agonists, ipratropium,glucocorticoids, methylxanthines, sodium channel blockers, opioidreceptor agonists, cannabinoid receptor agonists, cannabis, calciumchannel blockers, membrane stabilizers, acetaminophen, ibuprofen,aspirin, leukotriene inhibitors, any combinations thereof, or others. Inother embodiments, the compound(s) of the present invention may beuseful in combination therapy with other treatments for pain that do notcomprise an additional agent, e.g., meditation therapy, massage therapy,psychological therapy.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are useful for thetreatment of other somatosensory disturbances associated with ademyelinating disorder (e.g., tingling, numbness). Examples of suchagents include, but are not limited to, neurotrophin-3, antidepressants,anti-epileptic drugs, synthetic cannabinoids, pregabalin, gabapentin,sodium valproate, among others. In other embodiments, the compound(s) ofthe present invention may be useful in combination therapy with othertreatments for somatosensory disturbances that do not comprise anadditional agent, e.g., transcutaneous electrical nerve stimulation,meditation therapy, psychological therapy.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are useful for thetreatment of motor dysfunction associated with a demyelinating disorder.Examples of such agents include, but are not limited to,gamma-aminobutyric acid analogs, gamma-aminobutyric acid receptoragonists, benzodiazepines (e.g., clonazepam, carbamazepine), musclerelaxants, baclofen, anticholinergics, anticonvulsants (e.g., sodiumvalproate, phenytoin), anti-Parkinsonian agents (e.g., levadopa),antipsychotic agents (e.g., risperidone). In other embodiments, thecompound(s) of the present invention may be useful in combinationtherapy with other treatments for motor dysfunction that do not comprisean additional agent, e.g., chiropractic neurology, motor training, e.g.,rehearsal by eye movement, deep brain stimulation.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are useful for thetreatment of fatigue symptoms that may be associated with ademyelinating disorder. Examples of such agents include, but are notlimited to, iron supplements for anemia, medications and/or machines tohelp sleep apnea, sleeping pills, medications to control blood sugarlevels, medications to regulate thyroid function, antibiotics, vitamins,nutraceuticals (e.g., gingko biloba), antidepressants. In otherembodiments, the compound(s) of the present invention may be useful incombination therapy with other treatments for fatigue that do notcomprise an additional agent, e.g., sleep therapy, meditation therapy,diet or exercise changes, psychological therapy.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are useful for thetreatment of depression or other mood disorders that may be associatedwith a demyelinating disorder. Examples of such agents include, but arenot limited to, tricyclic antidepressants, tetracyclic antidepressants,selective serotonin reuptake inhibitors, serotonin/norepinephrinereuptake inhibitors, monoamine oxidase inhibitors, opioids (e.g.,buprenorphine), amphetamines, antipsychotics, benzodiazepines,nutraceuticals (e.g., St. John's Wort extract, Salvia elegans, Salviasclarea), tryptophan, omega 3 fatty acids, among others. In otherembodiments, the compound(s) of the present invention may be useful incombination therapy with other treatments for fatigue that do notcomprise an additional agent, e.g., sleep therapy, meditation therapy,diet or exercise changes, psychological therapy, electroconvulsivetherapy.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are useful for thetreatment of cognitive dysfunction that may be associated with ademyelinating disorder. Examples of such agents include, but are notlimited to, IFN-β 1a, IFN-β 1b, physostigmine, acetylcholinesteraseinhibitors (e.g., donepezil), nutraceuticals (e.g., gingko biloba),among others. In other embodiments, the compound(s) of the presentinvention may be useful in combination therapy with other treatments forcognitive dysfunction that do not comprise an additional agent, e.g.,cognitive retraining, psychological intervention, attention-trainingtasks, etc.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other agents that are useful for thetreatment of sexual dysfunction that may be associated with ademyelinating disorder. Examples of such agents include, but are notlimited to, vasoactive agents used for the treatment of erectiledysfunction (e.g., phenoxybenzamine phentolaimine, papverine,prostaglandin E1, vasoactive intestinal polypeptide, cyclic guanosine3′,5′,-monophosphate diesterase inhibitors such as sildenafil (Viagra),Tadalafil, Vardenafil, testosterone therapy), agents used for thetreatment of female sexual dysfunction (e.g., estrogen therapy, androgentherapy). In other embodiments, the compound(s) of the present inventionmay be useful in combination therapy with other treatments for sexualdysfunction that do not comprise an additional agent, e.g., changinglifestyle habits, meditation, yoga, psychological counseling, etc.

In some embodiments, the compound(s) of the present invention may beuseful in combination therapy with other treatments useful for themanagement of bladder or bowel dysfunction that may be associated with ademyelinating disorder. Examples of such treatments include: functionalelectrical stimulation, which may comprise implantation of an electricalstimulator that regulates the coordinated activation of muscles/nervescontrolling the bladder or bowel sphincter.

Encompassed in aspects of the present invention is the use of atherapeutically effective amount of a compound of Formula I and/or IIand one or more additional agents to be used in combination therapy.Also included in the subject methods is the use of a sub-therapeuticamount of a compound of Formula I and/or II and/or a sub-therapeuticamount of one or more additional agents. The individual components ofthe combination, though present in sub-therapeutic amounts, maysynergistically yield an efficacious effect and/or reduced adverseeffects in an intended application.

In addition, the compounds described herein may also be used incombination with other treatments not involving administration of anadditional agent, that may provide additional or synergistic benefit tothe patient. In one non-limiting example, subjects may experienceimproved therapeutic and/or prophylactic benefit in the methodsdescribed herein, wherein pharmaceutical composition of a compounddisclosed herein and/or combinations with other therapeutics arecombined with psychiatric or psychological therapy sessions (e.g., talktherapy sessions).

EXAMPLES Example 1

guanabenz protects differentiating rat oligodendrocyte precursor cells(drOPCs) against interferon-γ induced cell death (FIG. 2A). Interferon-γis an inflammatory cytokine that is thought to contribute to multiplesclerosis exacerbations, therefore, treatment of cultured cells of theoligodendrocyte lineage with interferon-γ provides a useful cellularmodel of MS-related inflammation. drOPCs were isolated and purifiedto >95% homogeneity from 6-7 day old Sprague-Dawley rat cortices byimmunopanning as previously described (Chan et al. 2004 Neuron 43:2;183-191). Briefly, rat cortices were extracted, diced, and digested withpapain at 37° C. Cells were then triturated and resuspended in panningbuffer containing insulin, then sequentially immunopanned at roomtemperature on three plates containing Ran-2, GalC, and O4 antibodiesfrom hybridomal supernatant. The remaining O4⁺GalC⁻ OPCs were removedfrom the plates with trypsin, resuspended in growth media, and seeded at37° C. onto pDL-coated flasks to facilitate proliferation. Oncesufficient numbers were reached, OPCs were split, plated indifferentiation media at 200,000 cells/well, and allowed todifferentiate for 24 hours. Differentiation media was then removed andreplaced with treatment media (containing IFN-γ (EMD Chemicals,Gibbstown, N.J.) and/or guanabenz (MP Biomedicals, Solon, Ohio)) whichwas refreshed after 24 hours. Following 48 hours of treatment, media wasremoved and the cells were incubated in FDA/PI(fluorescein-diacetate/propidium iodide) for 3 minutes, rinsed with1×PBS, and imaged for live/dead cell quantification. In living cells,non-fluorescent FDA is converted into fluorescein, giving off a brightgreen fluorescence. PI is membrane impermeable and therefore excludedfrom live cells. Therefore, green cells indicate viable cells and redcells indicate non-viable cells. Both control drOPCs not exposed toIFN-γ and 5.0 μM guanabenz-alone treated samples exhibit similar numbersof green and red cells, indicating that guanabenz alone does not impactcell viability (A,B). Treatment with 200 U/ml IFN-γ alone reduced thenumber of green cells and increased the number of red cells (C).Co-application of 1.25 μM and 2.5 μM guanabenz partially restoredviability (D, E), while 5.0 μM guanabenz increased the green/red cellratio back to control levels (F), indicating guanabenz protects cellsagainst IFN-γ induced cell death. FIG. 2B illustrates the quantitationof the experiment in FIG. 2a . Both untreated control and guanabenztreated samples exhibit comparable percent survival. Treatment withIFN-γ alone caused a roughly 40-50% decrease in cell survival.Co-application of 1.25 and 2.5 μM guanabenz increased cell survival byroughly 25% compared to IFN-γ alone samples, and 5.0 μM guanabenzincreased survival almost to control levels. Data is presented as themean+/−SEM for an N=3/group. (*p<0.05, **p<0.005)

Example 2

guanabenz protects differentiating rat oligodendrocyte precursor cells(drOPCs) against interferon-γ induced apoptosis (FIG. 3A). Cells wereisolated as described previously and plated in chamber slides at 20,000cells/chamber. Cells were then treated with the same IFN-γ and guanabenzprotocol as described previously, then subjected to a TUNEL assay.Treatment with IFN-γ alone caused a significant increase in apoptosis(as indicated by the increased numbers of green fluorescent cells), butco-treatment with 5.0 μM guanabenz reduced IFN-γ induced apoptosis.

FIG. 3B illustrates the quantitation of the experiment in FIG. 3a .Treatment with IFN-γ alone caused a significant (2-3 fold) increase inapoptosis compared to control cells, but co-treatment with 5.0 μMguanabenz reduced IFN-γ induced apoptosis by about 87.5% compared toIFN-γ alone. Error bars represent mean±SEM for an N=3/group (*p<0.05,**p<0.005, ***p<0.0005)

Example 3

immunohistochemical analysis of myelin basic protein and toluidine bluestaining demonstrates that guanabenz preserves myelin fibers andneuronal cytoarchitecture in ex vivo cultured cerebellar slices (FIG.4). Briefly, cerebellar sections were prepared from 6 day oldSprague-Dawley rat pups. Whole brains were extracted and embedded in 2%agarose in 1×PBS before cerebellar sections, including brain stem, weresectioned at a thickness of 300 μm via vibratome. Two to three sectionswere collected per well and grown in growth media with Fungizone.Sections were then switched to heat inactivated growth media and treatedwith 100 U/ml IFN-γ and or guanabenz for seven days, with half thevolume of media, including treatments, refreshed daily. Sections werethen stained for myelin-basic protein (A-E) and toluidine blue (F-H),used to visualize myelin and neuronal cytoarchitecture. Untreatedsections exhibit long, intact myelin fibers, while IFN-γ treatedsections exhibit fragmented structure, indicative of degenerated myelinfibers and axonal degeneration. Co-application of 2.5 μM, 5.0 μM, and10.0 μM guanabenz restored intact, long myelin fiber and axonalstructure in IFN-γ-treated sections.

Example 4

guanabenz delays symptom onset in an in vivo mouse model of chronicprogressive multiple sclerosis (FIGS. 5A and 5B). Subcutaneousinjections of 200 μg myelin oligodendrocyte glycoprotein (MOG) 35-55peptide emulsified in complete Freund's adjuvant (BD Biosciences, SanJose, Calif.) supplemented with 200 μg of Mycobacterium tuberculosis(strain H37Ra; BD Biosciences) were injected into the lower flanks of 8week old female C57BL/6J mice (Jackson Laboratory, Bar Harbor, Me.). Twointraperitoneal (IP) injections of 400 ng pertussis toxin each (ListBiological Laboratories, Denver, Colo.) were administered 0 and 48 hlater. Mice were monitored for clinical symptoms beginningpost-immunization day 7 (PID 7) and scored daily (0=healthy, 1=flaccidtail, 2=ataxia and/or paresis of hindlimbs, 3=paralysis of hindlimbsand/or paresis of forelimbs, 4=tetraparalysis, 5=moribund or death).Mice were treated intraperitoneally with guanabenz or vehicle (sterile0.9% NaCl) daily beginning PID 7. All protocols were approved by theUniversity of Chicago Institutional Animal Care and Use Committee.Vehicle-treated subjects exhibited rapid onset of symptoms (FIG. 5A).All guanabenz doses resulted in a significant delay in symptom onset(FIG. 5A). Note: bright red dots indicate average age of onset.N=14-15/group. (**p<0.005) FIG. 5B depicts quantitation of EAE onsetfrom the experiment described in FIG. 5a . Vehicle administered subjectsexhibited an average symptom onset of about 12 days. By contrast,treatment with 4 mg/kg/day guanabenz delayed symptom onset 19 days, 8mg/kg/day guanabenz delayed symptom onset to about 22 days, and 16mg/kg/day guanabenz delayed symptom onset to about 25 days. Error barsrepresent mean±SD for an N=14-15/group. (***p≤0.0005).

Example 5

guanabenz reduces symptom severity in an in vivo mouse model of chronicmultiple sclerosis (FIG. 6). Data from Example 4 were rearranged suchthat clinical scores were plotted as a function of time, as measured bydays post symptom onset. Vehicle-treated subjects exhibited clinicalscores of about 3.0. Neither 4 mg/kg/day nor 16 mg/kg/day caused asignificant reduction in clinical score, however, 8 mg/kg/day guanabenzsignificantly reduced symptom severity. Note: red bar indicates daysduring which *p≤0.05 compared to vehicle for the 8 mg/kg/day group.

Example 6

guanabenz reduces the severity of symptom relapse in an in vivo mousemodel of remitting/relapsing MS (FIG. 7A). Briefly, subcutaneousinjections of 50 μg proteolipid protein (PLP) 139-151 peptide emulsifiedin complete Freund's adjuvant (BD Biosciences) supplemented with 200 μgof Mycobacterium tuberculosis (strain H37Ra; BD Biosciences) wereinjected into the lower flanks of 8 week old female SJL mice (HarlanLaboratories, Indianapolis, Ind.). Mice were monitored for clinicalsymptoms beginning post-immunization day 7 (PID 7) and scored daily asdescribed above. Animals that did not achieve an acute phase (5 out of50) were removed from the study. Mice were then treated IP with 8 mg/kgguanabenz or vehicle (sterile 0.9% NaCl) daily at the beginning ofremission, defined as the second sequential day of reduced clinicalscore after the peak score of the acute phase. Mice that did not undergoa relapse phase (9 out of 22 vehicle-treated, 9 out of 23guanabenz-treated) were removed from the study. Animals treated with 8mg/kg guanabenz exhibited an average decrease in relapse severity ofabout 50%. Error bars represent mean±SEM for an N=13-14/group.

Example 7

higher doses of guanabenz also reduce symptom scores during theremission phase in an in vivo mouse model of remitting/relapsing MS(FIG. 7B). Briefly, SJL mice were exposed to PLP₁₃₉₋₁₅₁, monitored forclinical scores daily, and administered 16 mg/kg/day guanabenz asdescribed herein. Upon onset of remission (defined as reduction ofclinical score for at least two days after the peak score from thefirst, acute phase), animals were given daily IP injections of vehicleor guanabenz. Animals that exhibit relapse were assessed for severity ofclinical scores during relapse. Animals treated with 16 mg/kg guanabenzexhibited an average decrease in relapse severity of about 40%.Furthermore, animals treated with 16 mg/kg/day guanabenz also exhibiteda greater decrease in symptom severity during the remission phase,compared to vehicle-treated animals. N=11-13/group.

Example 8

guanabenz doses used in the in vivo and in vitro studies describedherein are comparable to doses used in humans (FIGS. 8, 9). C57Bl/6Jfemales immunized with MOG₃₅₋₅₅-specific EAE as described above weretreated daily IP with vehicle or 4, 8, or 16 mg/kg guanabenz diluted insterile 0.9% NaCl from PID 7 to around PID 35. Mice were then given afinal injection before serum was collected 2, 4, or 12 hours later viasubmandibular bleed. Immediately following blood draw, mice wereperfused with 0.9% NaCl and their brains collected and weighed. Sampleswere then outsourced for analysis of guanabenz concentrations byHPLC-MS/MS. FIG. 8 depicts guanabenz concentrations in blood serum (A)and brain (B). Though guanabenz appears to be quickly cleared in bothserum and brain in the mouse, the effective dose of 8 mg/kg appears tomaintain a concentration of ˜50 ng/ml in serum up to 4 hours aftertreatment (A). This concentration is comparable to that seen in humanplasma up to 36 hours after patients were given a single typical dose of32 mg of guanabenz (Meachem 1980, Clin Pharmacol Ther). This findingindicates that the dose found to be effective in alleviating anddelaying onset of EAE symptoms in mice is comparable to a dosage that isknown to be well-tolerated in humans. Guanabenz concentrations in braintissue are roughly 16-20× higher than those in serum (B). Datarepresents mean±SD. FIG. 9 depicts the same data, with brain and serumconcentrations expressed as μM values. Data depicted in FIGS. 2-3demonstrated that, compared to IFN-γ-challenged controls, 2.5-5.0 μMguanabenz in cell culture and 2.5-10 μM guanabenz in cerebellar sliceculture could significantly increase cell survival and inhibithypomyelination, respectively. Comparison of these concentrations tothose found in the serum (A) and particularly the brain tissue (B) ofmice treated with guanabenz indicate that the effective in vivo and invitro doses used are comparable. Data represents mean±SD.

Example 9

guanabenz reduces inflammatory immune responses to myelin proteins in anin vivo mouse model of EAE (FIG. 10). Mice immunized withMOG₃₅₋₅₅-specific EAE (as described herein) were treated with vehicle or8 mg/kg guanabenz daily IP from PID 7 to PID 19. On PID 17, mice werechallenged via a single injection of 10 μg of MOG₃₅₋₅₅ peptidesolubilized in 10 μl of 1×PBS in the dorsal surface of the left ear and10 μg of PLP₁₇₈₋₁₉₁ peptide in 10 μl 1×PBS in the right ear. Responsewas assessed 48 hours later by measuring ear thickness. 8 mg/kgguanabenz decreased mean ear swelling to about one-third ofvehicle-administered subjects. N=8-9/group. (**p<0.005, ***p<0.0005)

Example 10

co-administration of guanabenz and interferon delays onset of clinicalsymptoms in chronic EAE mice to a greater extent than either guanabenzalone or interferon alone (FIG. 11). Eight week old C57BL/6J female micewere immunized with MOG₃₅₋₅₅/CFA to induce chronic EAE. Vehicle(saline), guanabenz (8 mg/kg), interferon-beta (5,000 U/day/mouse) orcombined guanabenz/interferon-beta treatments were administered dailybeginning post immunization day 7. Mice were scored daily for clinicalsymptoms (0=healthy, 1=flaccid tail, 2=ataxia and/or paresis ofhindlimbs, 3=paralysis of hindlimbs and/or paresis of forelimbs,4=tetraparalysis, 5=moribund or death). Treatment with interferon alonedid not significantly delay onset of clinical symptoms. Onset ofclinical symptoms began at approximately day 12 in saline-treated miceand at approximately day 13 in mice treated with interferon alone. Onsetof clinical symptoms began at approximately day 18 in mice treated withguanabenz alone. Onset of clinical symptoms was further delayed in micetreated with guanabenz and interferon, beginning at approximately day 22for the co-administered mice. These results indicate a synergisticeffect of guanabenz and interferon for delaying onset of clinicalsymptoms of a demyelinating disorder. While preferred embodiments of thepresent invention have been shown and described herein, it will beobvious to those skilled in the art that such embodiments are providedby way of example only. Numerous variations, changes, and substitutionswill now occur to those skilled in the art without departing from theinvention. It should be understood that various alternatives to theembodiments of the invention described herein may be employed inpracticing the invention. It is intended that the following claimsdefine the scope of the invention and that methods and structures withinthe scope of these claims and their equivalents be covered thereby.

What is claimed is:
 1. A method of treating a demyelinating disorder ina subject in need thereof, comprising the step of administering to thesubject in need thereof an effective amount of a compound of Formula I:

or a derivative or pharmaceutically acceptable salt thereof, wherein R₁,R₂, R₃, R₄, and R₅ are independently hydrogen, deuterium, halogen,haloalkyl, alkyl, alkoxy, hydroxyl, aryl, or aryloxy, thereby treating ademyelinating disorder in said subject.
 2. The method of claim 1,wherein R₁ and R₅ are not hydrogen and/or wherein R₁ and R₅ are halogen.3. The method of claim 1, wherein the compound of Formula I is ofFormula II or pharmaceutically acceptable salt thereof:


4. The method of claim 1, wherein the demyelinating disorder is selectedfrom the group consisting of: multiple sclerosis, acute disseminatedencephalomyelitis, periventricular leukomalacia, periventricular whitematter injury, Tabes Dorsalis, Devic's disease, optic neuritis,progressive multifocal leukoencephalopathy, transverse myelitis, chronicinflammatory demyelinating polyneuropathy, anti-MAG peripheralneuropathy, adrenoleukodystrophy, adrenomyeloneuropathy, Guillain-BarréSyndrome, central pontine myelinolysis, diffuse white matter injury,leukodystrophy, and Charcot Marie Tooth Disease.
 5. The method of claim1, wherein treating comprises treating a symptom of a demyelinatingdisorder, wherein the symptom is selected from the group consisting of:fatigue, somatosensory dysfunction, tingling, pain, numbness, balanceproblems, problems with walking, changes in vision, depression,emotional changes, mood swings, impaired cognition, muscle dysfunction,impaired muscle coordination, sexual impairment, speech impairment,swallowing impairment, bladder dysfunction, bowel dysfunction.
 6. Themethod of claim 1, wherein about 1 mg to about 64 mg of the compound isadministered to the subject.
 7. The method of claim 1, wherein thecompound is administered orally.
 8. The method of claim 1, wherein theadministration of the compound is combined with one or more ofglatiramer acetate, dimethyl fumerate (BG-12), fingolimod (FTY720),interferon beta-1a, interferon beta-1b, mitoxantrone, natalizumab,dalfampridine, teriflunomide, and/or daclizumab.
 9. The method of claim1, wherein the compound of Formula I is guanabenz.
 10. The method ofclaim 9, wherein about 64 mg/day of said guanabenz is administered tosaid subject.
 11. The method of claim 1, wherein the compound of FormulaI is:

or a pharmaceutically acceptable salt thereof.
 12. The method of claim1, wherein the size and/or number of white matter plaques and/or lesionsis reduced.
 13. The method of claim 1, wherein treating comprisesprotecting brain cells from inflammation.
 14. The method of claim 1,wherein the administration of the compound is combined with glatirameracetate.
 15. A method of treating multiple sclerosis in a subject inneed thereof, comprising the step of administering to the subject inneed thereof an effective amount of

or a pharmaceutically acceptable salt thereof.
 16. A method forinhibiting hypomyelination in a subject in need thereof, comprising thestep of administering to the subject in need thereof an effective amountof

or a pharmaceutically acceptable salt thereof.
 17. The method of claim16, wherein the subject has or has been diagnosed with multiplesclerosis.